Formulations containing clopidogrel and sulfoalkyl ether cyclodextrin and methods of use

ABSTRACT

The present invention provides compositions containing clopidogrel, present as a free base or a pharmaceutically acceptable salt thereof, and sulfoalkyl ether cyclodextrin (SAE-CD). The compositions can be liquid, suspension or solid compositions. They can be adapted for oral, peroral or parenteral administration. The SAE-CD serves to aid in dissolution and stabilization of the clopidogrel in aqueous media. The stability of clopidogrel against hydrolytic degradation, thermal degradation, and photolytic degradation are improved. SAE-CD provides improved results over other cyclodextrin derivatives. The SAE-CD-containing composition of clopidogrel can be provided in liquid form, solid form or as a reconstitutable powder. Both ready-to-use and concentrated liquid compositions can be prepared. The liquid composition is optionally available as a clear solution. The compositions herein can be administered perorally or parenterally and provide substantial pharmacokinetic, pharmacodynamic and/or therapeutic advantages over a tablet composition administered perorally and excluding SAE-CD.

INCORPORATION BY REFERENCE TO ANY PRIORITY APPLICATIONS

Any and all applications for which a foreign or domestic priority claimis identified in the Application Data Sheet as filed with the presentapplication are hereby incorporated by reference under 37 CFR 1.57.

BACKGROUND OF THE INVENTION

1. Field of the Invention

The present invention relates to composition containing clopidogrel anda sulfoalkyl ether cyclodextrin and to the uses thereof in the treatmentof disorders and diseases that are therapeutically responsive toclopidogrel and to other methods of use thereof.

2. Description of the Related Art

Clopidogrel bisulfate, methyl(+)-(S)-α-(2-chlorophenyl)-6,7-dihydrothieno[3,2-c]pyridine-5(4H)-acetatesulfate (1:1), is an inhibitor of ADP-induced platelet aggregationacting by direct inhibition of adenosine diphosphate (ADP) binding toits receptor and of the subsequent ADP-mediated activation of theglycoprotein GPIIb/IIIa complex. Clopidogrel selectively inhibits thebinding of adenosine diphosphate (ADP) to its platelet receptor and thesubsequent ADP-mediated activation of the glycoprotein GPIIb/IIIacomplex, thereby inhibiting platelet aggregation. Biotransformation ofclopidogrel is necessary to produce inhibition of platelet aggregation.An active metabolite responsible for the activity of the drug has beenisolated (Pereillo et al., Drug Metab. Disposition (2002), 30(11),1288-1295). Clopidogrel also inhibits platelet aggregation induced byagonists other than ADP by blocking the amplification of plateletactivation by released ADP. Clopidogrel does not inhibitphosphodiesterase activity.

Clopidogrel bisulfate is a white to off-white powder. It is practicallyinsoluble in water at neutral pH but freely soluble at pH 1. U.S. Pat.No. 4,847,265 discloses the dextro-rotary form of clopidogrel. U.S. Pat.No. 7,074,928, No. 6767,913, No. 6,504,030, No. 6,429,210, and No.6,504,030 disclose polymorphic forms of clopidogrel hydrogen sulfate.U.S. Pat. No. 6,858,734, No. 6,800,759, and No. 6,737,411 disclosevarious methods for preparing clopidogrel. Salt forms, polymorphs, andprocesses for preparation of clopidogrel are disclosed in U.S. PregrantPublications No. 20060154957, No. 20060100231, No. 20060074242, No.20060047121, No. 20060041136, No. 20050256152, No. 20050228012, No.20050203122, No. 20050143414, No. 20050049275, and No. 20050049226. PCTInternational Publication No. WO 03/66637 discloses a hydrochloride saltform of clopidogrel and the process for its preparation.

Clopidogrel is currently marketed in the United States under the tradename PLAVIX (Sanofi Aventis). It is supplied in tablet form containing75 mg equivalents of the clopidogrel base, even though the drug ispresent in the bisulfate salt form. It is also available in generictablet form (Apotex, Inc.). U.S. Pat. No. 6,914,141 discloses a tabletformulation containing clopidogrel bisulfate.

PLAVIX is an antiplatelet medication approved by the U.S. Food and DrugAdministration to reduce athero thrombotic events in 1) patients with ahistory of recent myocardial infarction (MI), recent stroke, orestablished peripheral arterial disease (PAD), and 2) patients withacute coronary syndrome (unstable angina/non-Q-wave MI) includingpatients who are to be managed medically and those who are to be managedwith percutaneous coronary intervention (percutaneous transluminalcoronary angioplasty (PTCA), stent, atherectomy, etc.) or coronaryartery bypass graft (CABG). PLAVIX is a prescription medication thatwhen taken daily can help reduce the risk of having a future heartattack or stroke. U.S. Pat. No. 5,576,328 discloses a method ofpreventing secondary ischemic events by the administration ofclopidogrel after onset of a primary ischemic event. U.S. Pat. No.6,071,514 discloses methods of treating thrombotic disorders byadministration of clopidogrel to subjects in need thereof. Von Beckerathet al. (Circulation (2005), 112, 2946-2950) disclose the results of aclinical study comparing the absorption, metabolism and antiplateleteffects of 300 mg, 600 mg and 900 mg loading doses of clopidogreladministered perorally prepared from crushed Plavix tablets. They reportthe maximum ADP-induced platelet aggregation occurs at about 4 hours.Plavix inhibited 5uM ADP-induced platelet aggregation by approximately23% (300 mg dose), 34% (600 mg dose), 39% (900 mg dose), with minimaldifferences in antiplatelet effect between the 600 and 900 mg doses.These time periods are substantially similar to those usually observedfollowing administration of whole PLAVIX tablets (Weerakody et al. Am.J. Cardiol. 2007:100:331-336).

In the clinical setting, PLAVIX tablets are administered orally prior tocertain interventional cardiology procedures such as percutaneouscoronary intervention (PCI) in order to decrease a patient's plateletaggregation and thereby reduce the risk of reocclusion or restenosisduring or after the procedure. The amount of clopidogrel administered toa patient is related to the projected time to the procedure for thatpatient. In general, the greater the amount of clopidogrel administeredthe shorter the time to reach the desired therapeutic effect (e.g.platelet aggregation inhibition). For an average dose (300 mgclopidogrel in a PLAVIX tablet), the typical time to reach the desiredtherapeutic effect (e.g. platelet aggregation inhibition) varies fromtwo to five hours. If there is an immediate need for the procedure (forexample, PCI in less than two to three hours), then a larger dose ofclopidogrel is administered than would be administered if the procedurewere to be performed after two, three or more hours. For example, apatient undergoing a procedure in 60-180 minutes post administration ofclopidogrel may be administered 600 mg. For a procedure starting 180minutes or longer after administration of clopidogrel, the patient maybe administered 300 mg. The reason this dosing strategy being employedis that there is an apparent in vivo dose saturation effect, wherebyincreasing the dose does not increase the overall efficacy of the drugbut merely increases the rate of therapeutic onset, i.e. increases therate at which a target inhibition of platelet aggregation is achievedpost-administration of the drug. The specific protocol used, and timesbefore and after which larger or small doses are used varies betweendifferent institutions, but most use larger doses when procedures areperformed more quickly post dosing.

However, it is undesirable to administer unnecessarily large amounts ofclopidogrel due to its toxicology profile. Clopidogrel side effectsinclude hemorrhage, stomach upset/pain, diarrhea, constipation,headache, dizziness, rash, flu-like symptoms, back/joint pain, unusuallylong bleeding, unusual or easy bruising/bleeding, black stools,vomiting, chest pain, swelling, depression, fever, persistent sorethroat, unusual weakness, vision changes, slurred speech, confusion,severe rash, itching, severe dizziness, or trouble breathing.

Moreover, if a patient presents with a severe cardiac event, e.g. ACS(acute coronary syndrome), there is a need to treat the patient asquickly as possible in order to minimize the risk of myocardial damage,which increases rapidly with time. A clinician must be able to diagnosea patient as rapidly as possible in order to determine the appropriateemergency medical treatment as rapidly as possible. The goal is to beable to treat the patient who would require PCI within ninety minutesafter presenting in a hospital with ACS, but this short time period isunrealistic or unreliable when using PLAVIX tablets. Moreover, whileclopidogrel is contraindicated in major invasive emergency surgicalprocedures, such as CABG, it is indicated for minimally invasiveemergency procedures, such as PCI. One standard treatment protocol atthis time is as follows: 1) determine whether the patient is presentingwith an ACS; 2) alert the catheter lab of incoming patient; 3) oraladministration of 300-600 mg of clopidogrel (as PLAVIX tablet(s)); 4)transport patient to catheter lab; 5) perform coronary angiogram; 6)determine if medical therapy alone, PCI or CABG is most appropriate; and7) if medical therapy alone is indicated treat the patient with longterm (chronic) clopidogrel therapy; or 8) if PCI is indicated, performPCI and maintain patient on long term (chronic) clopidogrel therapy; or8) if CABG is indicated, delay CABG for until platelet aggregationreturns to normal for the patient. If CABG is performed within 7 daysafter the patient has received a dose of clopidogrel, there is a highrisk of major bleeding, hemorrhage-related complications, andtransfusion requirements (Pickard et al. Pharmacotherapy (2008), 23,376-392). Unfortunately, if administration of PLAVIX tablets is delayeduntil after a determination that PCI instead or CABG is indicated, thereis an increased risk of reocclusion and restenosis to the patientundergoing PCI. In addition, it is difficult to administer an oraltablet to a sedated patient which is often the case in subjectsundergoing coronary angiography.

Accordingly, it would be highly beneficial to this field of therapy toprovide a formulation that provides a more rapid therapeutic onsetwithout requiring such excessive doses as are currently administered.

Clopidogrel can be taken with another drug to treat a disorder ordisease in a subject. Clopidogrel, when taken with aspirin, isrecommended for people who have been hospitalized with heart-relatedchest pain or had a certain type of heart attack—conditions doctors callacute coronary syndrome (ACS). U.S. Pat. No. 7,018,990 discloses thecombined administration of a factor Xa inhibitor with clopidogrel. U.S.Pat. No. 6,509,348 discloses the combined administration of anADP-receptor blocking antiplatelet drug and a thromboxane A2 receptorantagonist and a method for inhibiting thrombus formation with thecombination. U.S. Pat. No. 6,248,729 discloses the combinedadministration of an ADP-receptor blocking antiplatelet drug andantihypertensive drug for preventing a cerebral infarction. U.S. Pat.No. 5,989,578 discloses the combined administration of clopidogrel andan antithrombotic agent. The combined use of clopidogrel with otherdrugs is also disclosed in U.S. Pregrant Publications No. 20050043382and No. 20040067995, and in published articles by Wegert et al. (Int. J.Clin. Pharmacol. Ther. (2002), 40(4), 135-141) and Gurbel et al.(Circulation, (2005), 111(9):1153-1159).

Various U.S. patents and publications disclose formulations comprisingclopidogrel, for example, U.S. Pat. No. 6,923,988, No. 6,761,903, No.6,720,001, No. 6,569,463, No. 6,451,339, No. 6,429,210, No. 6,383,471,No. 6,294,192, and Publications No.20060223845, No.20060003002,No.20040115287, No.20030104048, No.20020032149.

Cyclodextrins are cyclic carbohydrates derived from starch. Theunmodified cyclodextrins differ by the number of glucopyranose unitsjoined together in the cylindrical structure. The parent cyclodextrinscontain 6, 7, or 8 glucopyranose units and are referred to as α-, β-,and γ-cyclodextrin respectively. Each cyclodextrin subunit has secondaryhydroxyl groups at the 2 and 3-positions and a primary hydroxyl group atthe 6-position. The cyclodextrins may be pictured as hollow truncatedcones with hydrophilic exterior surfaces and hydrophobic interiorcavities. In aqueous solutions, these hydrophobic cavities provide ahaven for hydrophobic organic compounds, which can fit all, or part oftheir structure into these cavities. This process, known as inclusioncomplexation, may result in increased apparent aqueous solubility andstability for the complexed drug; however, the degree of stabilizationwill vary from drug to drug. The complex is stabilized by hydrophobicinteractions and does not involve the formation of any covalent bonds.

Chemical modification of the parent cyclodextrins (usually at thehydroxyl moieties) has resulted in derivatives with sometimes improvedsafety while retaining or improving the complexation ability of thecyclodextrin. Of the numerous derivatized cyclodextrins prepared todate, only two appear to be commercially viable; the 2-hydroxypropylderivatives (HP-β-CD or HPCD), neutral molecules being commerciallydeveloped by Janssen and others, and the sulfoalkyl ether derivatives(SAE-β-CD or SAE-CD), being developed by CyDex Pharmaceuticals, Inc.

The SAE-CDs are a class of negatively charged cyclodextrins, which varyin the nature of the alkyl spacer, the salt form, the degree ofsubstitution and the starting parent cyclodextrin. The sodium salt ofthe sulfobutyl ether derivative of beta-cyclodextrin, with an average ofabout 7 substituents per cyclodextrin molecule (SBE7-β-CD), is beingcommercialized by CyDex Pharmaceuticals, Inc. (Kansas) as CAPTISOL®cyclodextrin.

The anionic sulfobutyl ether substituent dramatically improves theaqueous solubility of the parent cyclodextrin. Reversible, non-covalent,complexation of drugs with the CAPTISOL® cyclodextrin generally allowsfor increased solubility and stability of some drugs in aqueoussolutions. However, the improved properties of SAE-CD over HP-β-CD interms of binding to specific drugs are somewhat unpredictable. Manydrugs are known to bind better with SAE-CD, while others are known tobind better with HP-β-CD. Moreover, CAPTISOL® cyclodextrin is relativelynew, and its combined use with clopidogrel has not been evaluated orsuggested in the prior art.

Various patent references disclosing compositions containing differentsalt, amorphous, crystalline and/or polymorphic forms of clopidogrel,optionally in the presence of another drug, suggest that the clopidogrelmight be included in such compositions as a complex with a cyclodextrin.However, none of those references specifies or exemplifies SAE-CD.

Slovenian Patent No. SI 21748, which issued Oct. 31, 2005 to RudolfRucman (DIAGEN D. O. O.) discloses inclusion complexes of clopidogrel,as a free base or salt form, and cyclodextrins such as β-CD, γ-CD,methyl cyclodextrin and hydroxyalkyl cyclodextrin, the latter two beingpreferred. The patent also discloses the use of poly (vinyl pyrrolidone)having a molecular weight of 10,000-40,000 instead of cyclodextrin tosolubilize the clopidogrel.

Kolbe et al. (J. Inclusion Phenomena and Macrocyclic Chemistry (2002Dec), 44(1-4), pg. 183-184) disclose the formation of a complex ofdimethylcyclodextrin and clopidogrel base in a 1:1 molar ratio. Thecomplex precipitates from cold solution.

U.S. Application Publication No. 2004-0109888 to Pun et al. discloses apolymeric cyclodextrin material.

U.S. Application Publication No. 2005-0096296 to Fikstad et al.discloses a “pharmaceutical composition comprising: a therapeuticallyeffective amount of a drug; a solubilizer; and a release modulator;wherein the release of the drug and solubilizer are synchronized.”

U.S. Application Publication No. 2005-0276841 to Davis et al. disclosesa “sustained-release biodegradable polymeric drug-eluting fiber”,wherein the drug may be complexed with a cyclodextrin.

Clopidogrel is known to possess poor chemical stability in solution. Itsdegradation typically proceeds by a hydrolytic pathway whereby the esterform is converted to the carboxylic acid derivative. The stability ofclopidogrel toward hydrolysis is pH dependent having a t₉₀ of about 52.7days at pH 5.6 when stored at 37° C. temperature in 0.1 M phosphatebuffer (Drug Metab. Disposition (2000), 28(12), 1405-1410). A chemicallystable solution formulation of clopidogrel would be useful in the art.Clopidogrel is known to undergo chiral inversion in vivo and in vitro(Reist et al., Drug Metab. Dispos. (2000), 28(12), 1405-1410); however,the (R)-enantiomer of clopidogrel is devoid of antithrombotic activityand can provoke convulsions in animals.

Due to the toxicology profile of clopidogrel and its typicaladministration at excessively high doses in a clinical procedure roomenvironment, it would be desirable to provide a formulation that couldavoid administration of the unnecessarily large amounts of clopidogrel,while at the same time provide the desired increase in the rate oftherapeutic onset, i.e. a desired decrease in the time after drugadministration that it takes to achieve the desired reduction inplatelet aggregation.

SUMMARY OF THE INVENTION

The invention provides a pharmaceutical composition comprisingclopidogrel (or any pharmaceutically acceptable salt thereof),sulfoalkyl ether cyclodextrin (SAE-CD), and optionally one or morepharmaceutically acceptable excipients. The SAE-CD is primarilyresponsible for solubilizing and stabilizing clopidogrel when the twoare in the presence of an aqueous medium. The compositions of theinvention reduce the chemical degradation of clopidogrel in solution.They also reduce the rate of chiral inversion of (S)-clopidogrel to(R)-clopidogrel. Even though the salt form of clopidogrel can beincluded in the instant formulation, the formulation does notnecessarily require exposure of clopidogrel to strongly acidicconditions, since the free-base form, rather than the salt form, ofclopidogrel can be employed in the formulation.

The invention provides an aqueous (optionally clear) liquid formulationcomprising SAE-CD, clopidogrel, and an aqueous liquid carrier.

The formulation of the invention can be a single-dose or multi-doseformulation. The claimed formulation can also be self-preserved againstmicrobial proliferation when the SAE-CD is present in amounts sufficientto stop or reduce the rate of microbial growth once the formulation hasbecome contaminated with a microbe. The present formulation alsoimproves the photochemical and thermal stability of clopidogrel overother cyclodextrin-based formulations.

The present invention also provides a SAE-CD-based solution ofclopidogrel that is pharmaceutically stable and that does not requiredilution prior to administration.

In some embodiments, 1) the sulfoalkyl ether cyclodextrin is present inan amount sufficient to provide a clear solution; 2) the SAE-CD ispresent at a concentration of about 20-600 mg/ml, 50-500mg/ml or 100-400mg/ml (2-60%, 5-50% or 10-40% wt./vol, respectively); 3) the SAE-CD isSBEx-β-CD, wherein x is 6.0 to 7.1 or 6.5 to 7; 4) the SAE-CD isSBEx-γ-CD, wherein x is about 6 to 8; and/or 5) the SAE-CD is a compoundof the formula 1 (infra.) or a mixture of compounds thereof.

In some embodiments, 1) the clopidogrel is present in therapeuticallyeffective amounts; and/or 2) clopidogrel as free base equivalents ispresent at a concentration of about 1.5 to 20 mg/ml (about 4.7 to 62 mM)or about 0.15 to 1.5 mg/ml (about 0.47 to 4.7 mM).

The invention also includes embodiments wherein: 1) the molar ratio ofSAE-CD to clopidogrel is in the range of at least about 6:1 or about 6:1to about 8:1 when the pH of the formulation is about or greater thanabout 3.5; 2) the molar ratio of SAE-CD to clopidogrel is less thanabout 6:1 when the pH of the formulation is less than about 3.5; 3) theclopidogrel is present at a concentration of about 7.5 mg/ml or less inan aqueous solution comprising about 37% wt/v or less of SAE-CD at a pHof about 5.5; 4) the clopidogrel is present at a concentration of about0.5 mg/ml or less in an aqueous solution comprising about 2.5% wt/v orless of SAE-CD at a pH of about 5.5; 5) clopidogrel as free baseequivalents is present at a concentration of about 0.15 to 20 mg/ml(about 0.47 to 62 mM); and/or 6) clopidogrel as bisulfate saltequivalents is present at a concentration of about 0.2 to about 26 mg/ml(about 0.47 to about 62 mM), about 0.2 to about 2 mg/ml (about 0.47 toabout 4.7 mM), or about 2 to about 26 mg/ml (about 4.7 to about 62 mM).

In some embodiments, the formulation does not require dilution prior toadministration to a subject. In other embodiments; the liquidformulation is dilutable with an aqueous based diluent withoutprecipitation of the clopidogrel.

In some embodiments, the formulation further comprises a solubilizingagent, a flavoring agent, a sweetening agent, a viscosity inducingagent, an antioxidant, a buffering agent, an acidifying agent, acomplexation enhancing agent, a lyophilizing aid (for example, bulkingagents or stabilizing agents), an electrolyte, another therapeuticagent, an alkalizing agent, an antimicrobial agent, an antifungal agentor a combination thereof.

In general, the liquid formulation has improved photochemical stabilityand undergoes less photolytic degradation when exposed to fluorescentlight as compared to another liquid formulation containing a differentcyclodextrin or derivatized cyclodextrin.

In some embodiments, the formulation has improved chemical stability,such as improved stability against hydrolysis of clopidogrel, andundergoes less hydrolytic degradation of clopidogrel when exposed toaqueous conditions as compared to other formulations wherein the SAE-CDhas been replaced by equimolar amounts of another cyclodextrin, such asHP-β-CD.

The invention also provides a method for preparing an aqueous(optionally clear) liquid formulation from a reconstitutable solid, themethod comprising the steps of: providing a reconstitutable solidcomprising clopidogrel, SAE-CD and optionally at least one otherpharmaceutical excipient, wherein the solid is reconstitutable with anaqueous liquid, and the molar ratio of SAE-CD to clopidogrel is at leastabout 6:1, at least about 6:1 to 8:1, or at least about 8:1 when the pHof the aqueous liquid is greater than or equal to about 3.5; andreconstituting the solid with a sufficient amount of aqueous liquidcarrier sufficient to at least suspend the reconstitutable solid,thereby forming the aqueous (optionally clear) liquid formulation.

In some embodiments, the liquid formulation has been prepared byreconstitution of a reconstitutable solid comprising at least SAE-CD andclopidogrel with an aqueous solution, wherein the reconstitutable solidis as defined herein. Conversely, the liquid formulation can belyophilized or otherwise dehydrated to form a reconstitutable solid.

Some embodiments of the invention include those wherein: 1) the liquidformulation is a suspension; 2) the amount of liquid carrier added issufficient to render the liquid formulation clear; 3) the formulationhas a concentration of clopidogrel in the range from about 1.5 to 20mg/ml or about 0.15 to 1.5 mg/ml; 4) the pH of the formulationapproximates or is less than the pKa of clopidogrel; 5) the pH of theformulation approximates or is greater than the pKa of clopidogrel; 6)the pH of the formulation is in the range of about 4-8 for parenteral ororal delivery; 7) the pH of the formulation is in the range of about 1to 3, 1 to 4, 1 to 8, 4 to 8, or 4 to 6 for oral delivery; 8) the molarratio of SAE-CD to clopidogrel is less than about 6:1 when the pH of theformulation is less than about 3.5; 9) the molar ratio of SAE-CD toclopidogrel is in the range of at least about 6:1 or about 6:1 to 8:1when the pH of the formulation is about or greater than about 3.5;and/or 10) the molar ratio is at least about 7.25:1 or 7.3:1 when the pHof the formulation is about or greater than about 8; the molar ratio isat least about 6.5:1 or 6.6:1 when the pH of the formulation is about orgreater than about 5.5.

Some embodiments of the invention include those wherein: 1) the methodfurther comprises the step of mixing the reconstitutable solid andaqueous liquid carrier; and/or 2) after reconstitution, the liquidformulation is ready for administration to a subject without requiringfurther dilution.

The invention also provides solid dosage forms. Such dosage forms can beadministered orally, enterally, buccally, sublingually or by other knownmodes of administration for solid dosage forms. The solid dosage formscan include a tablet, capsule, powder, reconstitutable solid and othersuch dosage forms. Parenteral administration of the dosage form could beperformed after dissolution of the dosage form in an aqueous liquidcarrier. Orally administered solid oral dosage forms may require loweramounts of SAE-CD depending upon the targeted region of release in thegastrointestinal tract. For solid oral dosage forms releasing in thegastric region, the molar ratio of SAE-CD to clopidogrel can be lessthan 6 to 1, less than 5 to 1, less than 4 to 1, less than 3 to 1, lessthan 2 to 1, less than 1 to 1, less than 0.5 to 1, less than 0.25 to 1and/or at least 0.05 to 1. For solid oral dosage forms releasing in thepost-gastric region(s), the molar ratio of SAE-CD to clopidogrel can beat least 0.2 to 1, at least 0.5 to 1, at least 1 to 1, at least 2 to 1,at least 3 to 1, at least 4 to 1, at least 5 to 1, at least 6 to 1, atleast 7 to 1, at least 8 to 1, and/or at most 100:1, at most 75:1, atmost 50:1, at most 40:1, at most 35:1, at most 30:1, at most 20:1, atmost 15:1, at most 12:1 or at most 10:1.

Combinations of the various upper and lower limits to the molar ratio ofSAE-CD to clopidogrel, as set forth in this disclosure, can be used toprovide different embodiments of the invention.

The invention also provides a method of administering clopidogrelcomprising administering a ready-to-use liquid or a solid oral dosageformulation comprising a sulfoalkyl ether cyclodextrin and clopidogrelor a pharmaceutically acceptable salt thereof.

Some embodiments of the methods of the invention include thosewherein: 1) the liquid formulation is administered parenterally,enterally or perorally; 2) the method further comprises the step ofdiluting a concentrate, according to the invention, with an aqueousliquid carrier prior to administration, thereby providing theready-to-use liquid formulation; 3) the method further comprises thestep of forming the liquid formulation by mixing an aqueous liquidcarrier with a reconstitutable solid according to the invention; or 4)the liquid formulation is formulated as described herein.

The present invention also provides a method of treating, preventing orreducing the occurrence of a disease, disorder or condition having anetiology associated with platelet aggregation or of a disease, disorderor condition that is therapeutically responsive to clopidogrel therapy,the method comprising administering the formulation of the invention toa subject in need thereof. Some embodiments of the invention includethose wherein the thrombotic disease, disorder, or condition is selectedfrom the group consisting of myocardial infarction, stroke, vasculardeath in patients with established peripheral arterial disease (PAD),secondary ischemic events, acute coronary syndrome (unstableangina/non-Q-wave MI, heart attack, angina), transient ischemic attack,cerebrovascular disease cardiovascular disease, angina pectoris, deepvein thrombosis (DVT), pulmonary emboli (PE), sickle cell crisis, andcardiac arrhythmia.

The invention also provides a method of decreasing the time totherapeutic onset or the time required to reach the target therapeuticeffect provided by clopidogrel, comprising administering parenterally,to a subject in need thereof, a formulation according to the invention;or administering orally or enterally, to a subject in need thereof, aformulation according to the invention. The formulation of the inventionprovides a reduced time to therapeutic onset and/or to targettherapeutic effect as compared to a solid oral dosage form administeredorally. The formulation of the invention also permits administration ofa lower dose of clopidogrel to achieve a target therapeutic effect, e.g.target bleeding time or target inhibition of platelet aggregation, ascompared to administration of a reference solid oral dosage formexcluding SAE-CD to achieve the same target therapeutic effect.

The invention provides a method of increasing the bleeding time in asubject comprising administering to a subject in need thereof acomposition comprising: SAE-CD; and no more than 900 mg, 750 mg, 675 mg,600 mg, 450 mg, 375 mg, 300 mg, 225 mg, 200 mg, 150 mg, 100 mg, 75 mg,50 mg, 40 mg, 30 mg, 25 mg, 20 mg, 15 mg, 12.5 mg, 10 mg, 7.5 mg, 5 mg,2 mg, 1 mg, 0.75 mg, or 0.1 mg of clopidogrel, or in the range of 0.1 to900 mg, 0.1 to 100 mg, 100 to 300 mg, about 300 mg, 300 to 600 mg, 300to 900 mg, 600 to 900 mg, 50 to 600 mg, 75 to 600 mg, 150 to 600 mg, or200 to 450 mg of clopidogrel, whereby the bleeding time of the subjectincreases by at least 10%, at least 15%, at least 20%, at least 25%, atleast 30%, at least 40%, at least 50%, at least 75%, at least 100%, atleast 150%, at least 200%, at least 250%, at least 300%, at least 400%,at least 500%, at least 700%, at least 900%, or at least 1000% during aperiod of no more than 200 min, 150 min, 120 min, 100 min, 90 min, 75min, 60 min, 50 min, 45 min, 40 min, 30 min, 15 min, 10 min, 7.5 min, 5min, 2.5 minutes, or of no more than 1 min, or of at least 10 seconds,or of 10 sec to 120 min, 30 sec to 100 min, 30 sec to 90 min, 30 sec to60 min, 1 min to 60 min, 1 min to 45 min, 1 min to 30 min, 1 min to 20min, or 1 min to 15 min following administration of the formulation,wherein said increase is determined by comparison to the subject'sbleeding time prior to administration of the composition. The upperlimit to percentage increase in the bleeding time can be up to 10,000%,up to 9,000%, up to 7,500%, up to 5,000%, up to 4,000%, up to 2,500%, orup to 1,000%. The method can comprise daily or chronically administeringto a subject in need thereof for several days, a week, several weeks, amonth, several months, three to twelve months, or more than a year.

The invention also provides a method of increasing the bleeding time ina subject in need thereof just prior to the subject undergoing a medicalprocedure, the method comprising administering to the subject aformulation comprising SAE-CD and no more than 900 mg, 750 mg, 675 mg,600 mg, 450 mg, 375 mg, 300 mg, 225 mg, 200 mg, 150 mg, 100 mg, 75 mg,50 mg, 40 mg, 30 mg, 25 mg, 20 mg, 15 mg, 12.5 mg, 10 mg, 7.5 mg, 5 mg,2 mg, 1 mg, 0.75 mg, or 0.1 mg of clopidogrel, or in the range of 0.1 to900 mg, 0.1 to 100 mg, 100 to 300 mg, about 300 mg, 300 to 600 mg, 300to 900 mg, 600 to 900 mg, 50 to 600 mg, 75 to 600 mg, 150 to 600 mg, or200 to 450 mg of clopidogrel, wherein the formulation is administered nomore than 200, 150, 100, 75, 60, 50, 40, 30, 15, 10, 7.5, 5 or 2.5minutes prior to the procedure and the bleeding time of the subjectincreases by at least 10, 15, 20, 25, 30, 40, 50, 75, 100, 150, 200,250, 300, 400, 500, 700, 900, or 1000%. during that time. The method cancomprise acutely administering one or more doses to a subject in needthereof just prior to the subject undergoing a medical procedure. Themethod can also comprise acutely administering a single dose to asubject in need thereof just prior to the subject undergoing a medicalprocedure such a as an interventional or non-interventional procedure.

The invention provides a method of decreasing extent of (or potentialfor) platelet aggregation in the blood of a subject in need thereofcomprising administering to the subject a formulation comprising SAE-CDand no more than 900 mg, 750 mg, 675 mg, 600 mg, 450 mg, 375 mg, 300 mg,225 mg, 200 mg, 150 mg, 100 mg, 75 mg, 50 mg, 40 mg, 30 mg, 25 mg, 20mg, 15 mg, 12.5 mg, 10 mg, 7.5 mg, 5 mg, 2 mg, 1 mg, 0.75 mg, or 0.1 mgof clopidogrel, or in the range of 0.1 to 900 mg, 0.1 to 100 mg, 100 to300 mg, about 300 mg, 300 to 600 mg, 300 to 900 mg, 600 to 900 mg, 50 to600 mg, 75 to 600 mg, 150 to 600 mg, or 200 to 450 mg of clopidogrel,whereby the percentage of platelet aggregation of the subject decreasesby at least 5, 10, 15, 30, 40, 50, 60, 70, 80, 90, 96, 98, or 100%during a period of no more than 200 min, 150 min, 120 min, 100 min, 90min, 75 min, 60 min, 50 min, 45 min, 40 min, 30 min, 15 min, 10 min, 7.5min, 5 min, 2.5 minutes, or of no more than 1 min, or of at least 10seconds, or of 10 sec to 120 min, 30 sec to 100 min, 30 sec to 90 min,30 sec to 60 min, 1 min to 60 min, 1 min to 45 min, 1 min to 30 min, 1min to 20 min, or 1 min to 15 min following administration of theformulation.

The invention also provides a method of reducing the extent of (orpotential for) platelet aggregation by at least 5, 10, 15, 30, 40, 50,60, 70, 80, 90, 96, 98 or 100% in a subject in need thereof, the methodcomprising administering to the subject a formulation comprising SAE-CDand no more than 900, 750, 675, 600, 450, 375, 300, 225, 200, 150, 100,75, 50, 40, 30, 25, 20, 15, 12.5, 10, 7.5, 5, 2, 1, 0.75, or 0.1 mg ofclopidogrel; or 0.1 to 900 mg, 0.1 to 100 mg, 100 to 300 mg, about 300mg, 300 to 600 mg, 300 to 900 mg, 600 to 900 mg, 50 to 600 mg, 75 to 600mg, 150 to 600 mg, or 200 to 450 mg of clopidogrel on a daily basis.

The extent of or potential for platelet aggregation can be measured invivo or ex vivo (in vitro).

Even though it can be administered orally, the clear liquidformulation/dosage form of the invention is particularly suitable forparenteral administration. In particular parenteral administration maybe desirable when oral administration of the formulation may beundesirable, i.e. at occasions such as pre-procedure administration,post-procedure administration, and other such modes of administration,or when a subject is incapacitated or otherwise unable to receive anoral dose of the liquid or solid formulation. A procedure, in thisparticular case, is a medical procedure. The formulation of theinvention can also be administered orally.

The present invention also provides methods of preparing an SAE-CD-basedaqueous solution or solid dosage form of clopidogrel or apharmaceutically acceptable salt thereof.

The invention also provides a kit comprising a first pharmaceuticalcomposition comprising SAE-CD and a second pharmaceutical compositioncomprising clopidogrel or a pharmaceutically acceptable salt thereof.

The invention also provides a taste-masked oral formulation comprisingsulfoalkyl ether cyclodextrin, clopidogrel, a pharmaceuticallyacceptable carrier, optionally one or more other excipients.

Unless otherwise specified, the term clopidogrel includes the free baseor salt form, and the racemic form, optically pure (R) form, opticallypure (S) form, or optically enriched form of the compound. It alsoincludes the solid, suspended or dissolved forms of the compound. Thesalt form can be present as the hemihydrate, hydrate or anhydrous form.The salt can also be present in a pure crystalline form or polymorphicform.

The invention also provides a method of improving the stability ofclopidogrel in a formulation comprising clopidogrel, the methodcomprising adding SAE-CD to the formulation in an amount sufficient tocomplex with a substantial portion of the clopidogrel present in theformulation, thereby stabilizing the clopidogrel.

The invention comprises a method of stabilizing a liquid formulationcomprising clopidogrel and an aqueous liquid carrier, the methodcomprising: adding SAE-CD to the formulation in an amount sufficient tocomplex with a substantial portion of the clopidogrel, therebystabilizing the clopidogrel. The term “stabilizing” is taken to meanreducing the rate of, reducing the extent of, and/or inhibiting of thedegradation or chiral inversion of the clopidogrel in solution. Themolar ratio of SAE-CD to clopidogrel can vary, as described herein,according to the pH of the liquid formulation and the desired stabilityof the formulation. Generally, the greater the portion of clopidogrelthat is complexed with the SAE-CD, the greater the stabilization of theclopidogrel. Accordingly, the invention also provides a method ofstabilizing (S)-clopidogrel against chiral inversion to (R)-clopidogrel,the method comprising including a sulfoalkyl ether cyclodextrin in acomposition or formulation comprising an optically pure orenantiomerically enriched form of (S)-clopidogrel. By enantiomericallyenriched is meant that the composition comprises a greater amount of(S)-clopidogrel than of (R)-clopidogrel.

Since the SAE-CD stabilizes clopidogrel in solution, a formulation orcomposition of the invention is also a “stabilized formulation” or“stabilized composition”. A “stabilized formulation” or “stabilizedcomposition” possesses enhanced stability as compared to an otherwisesimilar formulation excluding the SAE-CD.

The invention also provides a formulation comprising SAE-CD,clopidogrel, a second therapeutic agent, and a pharmaceuticallyacceptable carrier. The second therapeutic agent can be a nonsteroidalantiinflamatory drug, anticoagulant, selective factor Xa inhibitor,direct thrombin inhibitor, antiplatelet agent, platelet aggregationinhibitor, glycoprotein Ilb/IIIa inhibitor, antisickling agent,hemorrheologic agent, thrombolytic agent, thrombolytic enzyme, tissueplasminogen activator, or combination thereof.

The invention also provides a method of treating a disease, condition ordisorder comprising administering to a subject in need thereof: atherapeutically effective amount of clopidogrel in a composition orformulation of the invention, and a therapeutically effective amount ofa second therapeutic agent, such as described herein. The secondtherapeutic agent may or may not be included in the same composition orformulation as the clopidogrel.

The invention also provides a method of identifying a responder ornon-responder subject as regards responsiveness to clopidogrel therapy,the method comprising: administering to the subject a compositioncomprising an expected therapeutically effective amount of clopidogrel,and determining the subject's responsiveness to clopidogrel within aperiod of about 10 sec to 120 min, 30 sec to 120 min, 30 sec to 100 min,30 sec to 90 min, 30 sec to 60 min, 1 min to 60 min, 1 min to 45 min, 1min to 30 min, 1 min to 20 min, or 1 min to 15 min, 1 min to 90 min, 5min to 60 min, 10 min to 60 min, 5 min to 45 min, 10 min to 45 min, 15min to 30 min, 5 min to 30 min, 5 min to 15 min, or 10 min to 20 min, orwithin less than about 10 seconds, less than about 1 min, less thanabout 2.5 min, less than about 5 min, less than about 7.5 min, less thanabout 10 min, less than about 15 min, less than about 20 min, less thanabout 30 min, less than about 120 min, less than about 100 min, lessthan about 90 min, less than about 75 min, less than about 60 min, lessthan about 50 min, less than about 45 min, or less than about 40 minafter administration of the composition to the subject.

The composition can be adapted for peroral or parenteral administration.The expected therapeutically effective amount of clopidogrel willgenerally be about 50 to 600 mg, 0.1 to 900 mg, 1 to 900 mg, 10 to 900mg, 0.1 to 100 mg, 25 to 750 mg, 50 to 600 mg, 75 to 600 mg, 75 to 500mg, 100 to 300 mg, 100 to 400 mg, 150 to 600 mg, or 200 to 450 mg, 200to 400 mg, 300 to 600 mg, 300 to 900 mg, 600 to 900 mg, about 0.1 mg,about 0.75 mg, about 1 mg, about 2 mg, about 5 mg, about 7.5 mg, about10 mg, about 12.5 mg, about 15 mg, about 20 mg, about 25 mg, about 30mg, about 40 mg, about 50 mg, about 75 mg, about 100 mg, about 150 mg,about 200 mg about 225 mg, about 300 mg, about 375 mg, about 450 mg,about 525 mg, about 600 mg, about 675 mg, about 750 mg, about 900 mg.

The step of determining can comprise: obtaining a sample of blood of thesubject; and determining the extent of platelet aggregation in thesubject's plasma by aggregometry, such as light transmittance orimpedance platelet aggregometry. The step of administering can compriseperoral or parenteral administration. The composition can furthercomprise an amount of SAE-CD sufficient to solubilize and/or stabilizethe clopidogrel.

The invention also provides a method of reducing the requiredtherapeutic dose in a responder subject in need of clopidogrel toachieve a target therapeutic effect therein, the method comprising:parenterally or perorally administering to the subject a pharmaceuticalcomposition comprising SAE-CD and a first therapeutically effectiveamount of clopidogrel, wherein the first therapeutically effectiveamount is at least 1.1-fold, at least 1.2-fold, at least 1.25-fold, atleast 1.5-fold, at least 2-fold, at least 3-fold, at least 4-fold, atleast 5-fold, at least 7-fold, at least 8-fold, at least 10-foldsmaller, at least 15-fold, at least 20-fold, about 1.1 to about 20-fold,about 1.2-fold to about 15-fold, about 1.25-fold to about 10-fold, about2-fold to about 10-fold, or about 3 fold to about 8-fold smaller than asecond therapeutically effective amount, which is the amount ofclopidogrel required to provide substantially the same therapeuticeffect when clopidogrel is administered to the subject perorally in areference solid pharmaceutical composition excluding SAE-CD. In someembodiments, the first therapeutically effective amount is no more than900 mg, 750 mg, 675 mg, 600 mg, 450 mg, 375 mg, 300 mg, 225 mg, 200 mg,150 mg, 100 mg, 75 mg, 50 mg, 40 mg, 30 mg, 25 mg, 20 mg, 15 mg, 12.5mg, 10 mg, 7.5 mg, 5 mg, 2 mg, 1 mg, 0.75 mg, or 0.1 mg of clopidogrel,or in the range of 0.1 to 900 mg, 0.1 to 100 mg, 50 mg to 600 mg, 100 to300 mg, about 300 mg, 300 to 600 mg, 300 to 900 mg, 600 to 900 mg, 50 to600 mg, 75 to 600 mg, 150 to 600 mg, or 200 to 450 mg and the secondtherapeutically effective amount is greater than or about 300 mg to 900mg for solid oral tablets, such as PLAVIX tablets.

The invention also provides a method of converting a subject that is anon-responder, in terms of oral administration of clopidogrel, to aresponder, the method comprising parenterally administering clopidogrelto a subject in need thereof, thereby providing a therapeutic responseto clopidogrel in the subject.

The invention also provides a method of escalating dose in a subject toachieve a target therapeutic effect in the subject, the methodcomprising: parenterally administering to the subject a first amount ofclopidogrel, which can be 10 mg to 600 mg, 50 mg to 600 mg, 50 mg to 300mg, about 50 mg, about 75 mg, about 100 mg, about 150 mg, about 225 mg,about 300 mg, about 375 mg, about 450 mg, about 525 mg, or about 600 mg;within a period of less than 120 min, less than about 100 min, less thanabout 90 min, less than about 75 min, less than about 60 min, less thanabout 45 min, less than about 30 min, less than about 20 min, less thanabout 15 min, less than about 10 min, less than about 5 min, or of 30sec to 120 min, 30 sec to 100 min, 1 min to 100 min, 1 min to 90 min, 5min to 90 min, 10 min to 75 min, 10 min to 60 min, 15 min to 60 min, 15min to 45 min, or 15 min to 30 min after the parenteral administration,determining the first corresponding therapeutic effect achieved in thesubject; if the extent of therapeutic effect achieved is less than thetarget therapeutic effect (for example, in terms of inhibition ofplatelet aggregation), parenterally administering to the subject asecond amount of clopidogrel, wherein the second amount is about0.5-fold, about 1-fold, about 1.25-fold, about 1.5-fold, or about 2-foldof the first amount, or the second amount is 0.1 mg to 1200 mg, 1 mg to1000 mg, 5 mg to 900 mg, 10 mg to 900 mg, 25 mg to 750 mg, 50 mg to 750mg, 50 mg to 600 mg, 0.1 mg to 100 mg, 1 mg to 75 mg, 100 mg to 300 mg,300 mg to 600 mg, or 600 mg to 1200 mg; determining the secondcorresponding therapeutic affect achieved in the subject afteradministration of the second amount; and if the second correspondingtherapeutic affect is less than the target therapeutic effect, repeatingthe steps of “parenterally administering to the subject a second amountof clopidogrel” and of “determining the second corresponding therapeuticaffect achieved” until the target therapeutic effect is achieved.

The step of determining can comprise: obtaining a sample of plasma ofthe subject; and determining the extent of platelet aggregation in thesubject's plasma by aggregometry. The step of obtaining can comprise:obtaining a sample of blood of the patient; and separating the plasmafrom the blood to form a plasma sample. The clopidogrel can be presentin a composition comprising SAE-CD. The invention also provides analternative method, wherein the clopidogrel is administered perorally.

The invention also provides a treatment protocol for a subjectpresenting with a cardiovascular condition, disease or disorder,optionally wherein the subject is initially not undergoing clopidogreltherapy the protocol comprising: a) determining whether or not thesubject requires interventional or non-interventional medical treatment;and b) if the subject requires minimally invasive interventional medicaltreatment, then administering to the subject a pharmaceuticalcomposition comprising SAE-CD and clopidogrel in an amount sufficient toprovide a target therapeutic effect in the subject within a period ofless than 120 min, less than about 100 min, less than about 90 min, lessthan about 75 min, less than about 60 min, less than about 45 min, lessthan about 30 min, less than about 20 min, less than about 15 min, lessthan about 10 min, less than about 5 min, or of 30 sec to 120 min, 30sec to 100 min, 1 min to 100 min, 1 min to 90 min, 5 min to 90 min, 10min to 75 min, 10 min to 60 min, 15 min to 60 min, 15 min to 45 min, or15 min to 30 min, and conducting the minimally invasive interventionalprocedure; or c) if the subject requires non-interventional medicaltreatment, then administering to the subject a pharmaceuticalcomposition comprising SAE-CD and clopidogrel in an amount sufficient toprovide a target therapeutic effect in the subject within a period of 10sec to 120 min, 30 sec to 120 min, 30 sec to 100 min, 30 sec to 90 min,30 sec to 60 min, 1 min to 60 min, 1 min to 45 min, 1 min to 30 min, 1min to 20 min, or 1 min to 15 min, 1 min to 90 min, 5 min to 60 min, 10min to 60 min, 5 min to 45 min, 10 min to 45 min, 15 min to 30 min, 5min to 30 min, 5 min to 15 min, or 10 min to 20 min, or within less thanabout 10 seconds, less than about 1 min, less than about 2.5 min, lessthan about 5 min, less than about 7.5 min, less than about 10 min, lessthan about 15 min, less than about 20 min, less than about 30 min, lessthan about 120 min, less than about 100 min, less than about 90 min,less than about 75 min, less than about 60 min, less than about 50 min,less than about 45 min, or less than about 40 min and providing to thesubject said non-interventional medical treatment; or d) if the subjectrequires invasive interventional medical treatment, then notadministering to the subject clopidogrel. In some embodiments, theminimally invasive interventional procedure is PCI. In some embodiments,the invasive interventional procedure is CABG. In some embodiments, theprotocol comprises: determining whether the subject is presenting withan ACS; optionally, alerting the catheter lab of incoming subject;optionally, transporting subject to the catheter lab; performingcoronary angiography on the subject; determining if PCI or CABG isindicated; and if PCI is indicated, parenterally administering a liquidcomposition comprising clopidogrel or perorally administering acomposition comprising clopidogrel and SAE-CD, performing PCI, andoptionally maintaining the subject on long term (chronic) clopidogreltherapy; or if CABG is indicated, performing the CABG, without prioradministration of clopidogrel. The method of the invention can includeadditional steps as needed, such as sedating the subject and/or anystep(s) indicated by an attending clinician/physician.

The invention provides a method of decreasing the time to peak or targettherapeutic effect in a responder subject administered clopidogrel, themethod comprising: administering to a subject in need thereof a firstcomposition comprising SAE-CD and a therapeutically effective amount ofclopidogrel sufficient to achieve a target therapeutic effect, wherebythe time to peak or target therapeutic effect achieved by administrationof the first composition is less than the time to peak or targettherapeutic effect achieved by similar administration of an otherwisesimilar reference composition, excluding SAE-CD, and comprisingsubstantially the same therapeutically effective amount of clopidogrel.In some embodiments, the time to peak or target therapeutic effect isreduced by at least 1.1-fold, at least 1.2-fold, at least 1.25-fold, atleast 1.5-fold, at least 2-fold, at least 3-fold, at least 4-fold, atleast 5-fold, at least 7-fold, at least 8-fold, at least 10-foldsmaller, at least 15-fold, at least 20-fold, at least 40-fold, at least50-fold, at least 75-fold, at least 100-fold, at least 120-fold, about1.1-fold to about 120-fold, about 2-fold to about 120-fold, 2-fold to100-fold, 2-fold to 75-fold, or 2-fold to 50-fold.

The invention also comprises combinations of the embodiments and aspectsof the invention as detailed herein. Accordingly, the invention alsoincludes combinations and sub-combinations of the individual elements ofthe embodiments or aspects of the invention as described herein. Otherfeatures, advantages and embodiments of the invention will becomeapparent to those skilled in the art by the following description,accompanying examples.

BRIEF DESCRIPTION OF THE DRAWINGS

The following drawings are part of the present specification and areincluded to further demonstrate certain aspects of the invention. Theinvention may be better understood by reference to one or more of thesedrawings in combination with the summary and detailed description of theembodiments presented herein.

FIG. 1 depicts data obtained from a room temperature phase solubilitystudy conducted with clopidogrel bisulfate and various different SAE-CDderivatives, underivatized cyclodextrins, and 2-hydroxypropyl-β-CD inwater at pH˜5.5.

FIG. 2 a depicts data obtained from a thermal stability study for thecombination of clopidogrel and either SBE-β-CD or HP-β-CD conducted in aphosphate buffer (0.1 and 0.2 M) at pH 5.5 and 60° C. prepared accordingto Example 19.

FIG. 2 b depicts data obtained from a thermal stability study for thecombination of clopidogrel and either SBE-β-CD or HP-β-CD conducted in aphosphate buffer (0.1 and 0.2 M) at pH 8.0 and 60° C. prepared accordingto Example 19.

FIG. 3 a depicts the chiral conversion data (from (S)-clopidogrel to(R)-clopidogrel) obtained from a thermal stability study for thecombination of clopidogrel and either SBE-β-CD or HP-β-CD conducted in aphosphate buffer (0.1 or 0.2 M) at pH 5.5 and 40° C. prepared accordingto Example 19.

FIG. 3 b depicts the chiral conversion (from (S) -clopidogrel to(R)-clopidogrel) data obtained from a thermal stability study for thecombination of clopidogrel and either SBE-β-CD or HP-β-CD conducted in aphosphate buffer (0.1 or 0.2 M) at pH 8.0 and 40° C. prepared accordingto Example 19.

FIG. 4 a depicts the chiral conversion (from (S)-clopidogrel to(R)-clopidogrel) data obtained from a fluorescent light stability studyfor the clopidogrel and either SBE-β-CD or HP-β-CD conducted in aphosphate buffer (0.1 or 0.2 M) at pH 5.5 and 29° C.

FIG. 4 b depicts the chiral conversion (from (S) -clopidogrel to(R)-clopidogrel) data obtained from a fluorescent light stability studyfor the clopidogrel and either SBE-β-CD or HP-β-CD conducted in aphosphate buffer (0.1 or 0.2 M) at pH 8.0 and 29° C.

FIG. 5 depicts data regarding the pH solubility profile of clopidogrelbisulfate at ambient temperature.

FIG. 6 a depicts a phase solubility diagram for clopidogrel bisulfate(mg/ml) in the presence of SAE-CD (wt./vol %) at 25° C. in water with pHadjusted to 5.5.

FIG. 6 b depicts a phase solubility diagram for clopidogrel bisulfate(M) in the presence of SAE-CD (M) at 25° C. in water with pH adjusted to5.5.

FIG. 7 depicts the results of a competitive binding assay between SAE-CDand clopidogrel salt in the presence of aspirin conducted according toExample 22

FIG. 8 depicts the results of an in vivo study to determine bleedingtime in mice following administration (parenteral and peroral) of aclear liquid formulation prepared according to Example 20. Theevaluation was conducted according to Example 21.

FIG. 9 depicts the results of the dissolution study of tablets made froma spray dried solid comprising SBE-β-CD and clopidogrel bisulfatesolution, wherein the weight ratio of SBE-β-CD to clopidogrel bisulfateis 250 mg to 98 mg, respectively, or made from a physical mixture ofSBE-β-CD and clopidogrel bisulfate using the same ratio. These werecompared to the marketed tablet PLA VIX®. The evaluation was conductedaccording to Example 23.

FIG. 10 a log-linear plot of mean plasma clopidogrel concentrationversus time after IV administration in patient cohorts receiving 0.1 to300 mg of clopidogrel in an aqueous liquid composition of the invention.

FIG. 11 a log-linear plot of mean plasma clopidogrel thiol activemetabolite concentration versus time after administration for thepatients of FIG. 10.

FIG. 12 a log-linear plot of mean plasma clopidogrel carboxylic acidmetabolite concentration versus time after administration for thepatients of FIG. 10.

FIG. 13 is a plot of the mean percent of platelet aggregation inhibitionversus time after IV administration in patient cohorts receiving 0.1 to300 mg of clopidogrel in an aqueous liquid composition of the invention.

FIG. 14 is a Kaplan-Meier plot for estimates of percentage of patientsachieving >/=15% platelet aggregation inhibition versus time afteradministration for the patients of FIG. 13.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENT

When prepared in ready-to-use (i.e., ready-to-administer) form, theliquid formulation of the invention does not require dilution prior toadministration. When present as a concentrate, the present formulationis also dilutable in a broad range of aqueous based diluents withoutformation of precipitate. The formulation can be an oral, peroral,enteral or parenteral formulation. As used herein and unless otherwisespecified, the term “clopidogrel” includes all neutral, free base, salt,crystalline, non-crystalline, amorphous, optically pure, opticallyenriched, racemic and/or polymorphic forms of the same. The clopidogrelcan be present in anhydrous or hydrated form prior to use in presentformulation. The salt of clopidogrel can be a pharmaceuticallyacceptable salt. The (S)-enantiomer of clopidogrel can be made accordingto U.S. Pat. No. 4,847,265. The (R)-enantiomer of clopidogrel can bemade according to French Patent No. FR 2769313. The racemic form ofclopidogrel can be made according to U.S. Pat. No. 4,529,596, and iscommercially available from Sigma-Aldrich (St. Louis, Mo.)

As used herein, “pharmaceutically acceptable salt” refers to derivativesof clopidogrel wherein the active agent is modified by reacting it withan acid as needed to form an ionically bound pair. Examples ofpharmaceutically acceptable salts include conventional non-toxic saltsor the quaternary ammonium salts of the parent compound formed, forexample, from non-toxic inorganic or organic acids. Suitable non-toxicsalts include those derived from inorganic acids such as hydrochloric,hydrobromic, sulfuric, sulfonic, sulfamic, phosphoric, nitric and othersknown to those of ordinary skill in the art. Other salts are preparedfrom organic acids such as amino acids, acetic, propionic, butyric,succinic, glycolic, gluconic, stearic, lactic, malic, tartaric, citric,ascorbic, pamoic, maleic, hydroxymaleic, phenylacetic, glutamic,benzoic, salicylic, sulfanilic, 2-acetoxybenzoic, fumaric,toluenesulfonic, methanesulfonic, ethanesulfonic, benzenesulfonic,oxalic, isethionic, and other acids known to those of ordinary skill inthe art. Lists of other suitable salts are found in Remington'sPharmaceutical Sciences, 17^(th) . ed., Mack Publishing Company, Easton,Pa., 1985 or 1995, the relevant disclosure of which is herebyincorporated by reference. Specific pharmaceutically acceptable salts ofclopidogrel include the bisulphate salt, hydrobromide, napsylate,benzensulfonate, prop ylsulf ate, perchlorate, naphthalenesulfonate,hydrochloride, isopropylsulfate, hydroiodide and mesylate.

As used herein, the term reconstitutable solid (reconstitutablecomposition) is taken to mean a solid capable of dissolution in anaqueous liquid medium to form a reconstituted liquid, wherein afterdissolution the liquid medium is suitable for administration. In oneembodiment, the reconstitutable solid forms a liquid formulation that isvisibly clear when the solid is mixed with aqueous carrier. Areconstitutable pharmaceutical formulation according to the presentinvention comprises clopidogrel, SAE-CD and optionally, at least oneother pharmaceutical excipient, wherein the molar ratio of SAE-CD toclopidogrel is as defined herein. A reconstitutable solid can beprepared by removal of the liquid medium from an aqueous liquid solutioncomprising SAE-CD and clopidogrel, and optionally other components toform the solid. The reconstitutable solid composition can comprise anadmixture of a solid SAE-CD and a clopidogrel-containing solid andoptionally at least one other pharmaceutical excipient, such that amajor portion of the clopidogrel is not complexed with the SAE-CD priorto reconstitution. Alternatively, the composition can comprise a solidmixture of SAE-CD, clopidogrel and optionally at least one otherpharmaceutical excipient, wherein a major portion of the clopidogrel iscomplexed with the SAE-CD prior to reconstitution. A reconstitutablesolid will generally comprise less than 8% wt. water. This compositioncan be reconstituted with an aqueous solution to form a liquidformulation containing clopidogrel and other agents that can beadministered to a subject. The liquid formulation used in thepreparation of a reconstitutable formulation may be prepared asdescribed herein for the diluted or concentrated liquid formulations. Itmay also be prepared to contain an SAE-CD and the clopidogrel atconcentrations greater than typically used in the liquid formulation ofthe invention, while maintaining the same SAE-CD to clopidogrel molarratio. Applicants note that any composition according to the inventioncan be dissolved or diluted with another liquid containing SAE-CD.

The reconstitutable composition can be prepared according to any of thefollowing processes. A liquid formulation of the invention is firstprepared, then a solid is formed by lyophilization (freeze-drying),spray drying, spray freeze-drying, vacuum-drying, antisolventprecipitation, various processes utilizing supercritical or nearsupercritical fluids, or other methods known to those of ordinary skillin the art of the liquid formulation to make a powder or a solidsuitable for reconstitution. As noted above, the reconstitutable solidcan be an admixture of the dry components, which is prepared byphysically blending the components in the absence of excess moisture,i.e. the moisture should be less than about 60% RH.

A reconstitutable solid can be a powder, glassy solid, porous solid,granulate, pellet, bead, compressed solid, particulate or lyophile.

As used in regards to an SAE-CD-containing composition or formulationaccording to the invention, the term dilutable refers to a liquidformulation containing SAE-CD and clopidogrel, wherein the formulationcan be further diluted with a clear aqueous liquid carrier at roomtemperature, e.g., ambient temperature such as a temperature of about20° -28° C., preferably without significant precipitation ofclopidogrel, i.e. if precipitation occurs it is less than or equal toabout 3% wt. of clopidogrel, while providing a final clear solution whendiluted to a clopidogrel concentration of about 0.15 to 10 mg/ml (freebase equivalents). When a dilutable SAE-CD and clopidogrel-containingformulation is diluted with a non-clear solution, the resulting mixturemay or may not be clear. A dilutable SAE-CD and clopidogrel-containingliquid can be diluted with another solution that does not containSAE-CD, and the resulting diluted solution will have a lowerconcentration of solubilized clopidogrel preferably without causingsignificant precipitation of clopidogrel.

Exemplary liquids for diluting an oral formulation of the inventioninclude commercially available beverages such as carbonated beverages,non-carbonated beverages, and juices. Exemplary carbonated beveragesinclude flavored and non-flavored sodas, wherein the flavor is a cola,lemon, lime, root beer, bubble gum, cherry, orange and other flavors ormixtures thereof. Exemplary juices include apple, lemon, lime, orange,grape, cherry, cranberry, grapefruit, strawberry, kiwi, raspberry,blueberry, blackberry, dewberry, tangerine, pineapple, watermelon,cantaloupe, ginger, guava, mango, papaya, plum, apricot, pear, peach,nectarine, pomegranate, and other juices or mixtures thereof.Accordingly, an SAE-CD and clopidogrel-containing solution that is notdilutable according to the invention will form a significant amount (>3%wt. of active agent) of precipitate when diluted with another solution.

It should be noted that a formulation that is not dilutable with wateralone at room temperature may be rendered dilutable with an aqueoussolution that contains SAE-CD as long as the final molar ratio ofclopidogrel to SAE-CD in the diluted solution is within the requiredrange as described herein. The invention therefore provides a method ofrendering dilutable a previously non-dilutable (as defined herein)clopidogrel-containing solution comprising the step of diluting thepreviously non-dilutable solution with a second solution containingSAE-CD such that the molar ratio of SAE-CD to clopidogrel in the dilutedsolution is as defined herein.

As used herein, a pharmaceutically acceptable liquid carrier is anyaqueous liquid medium used in the pharmaceutical sciences for dilutionor dissolution of parenteral, oral or peroral formulations, such aswater, aqueous buffer, aqueous organic solvent, and other liquidsdescribed herein or used in the pharmaceutical and/or food industry.

The formulation of the invention comprises clopidogrel and a sulfoalkylether cyclodextrin of the formula 1:

wherein:

n is 4, 5 or 6;

R₁, R₂, R3, R₄, R₅, RO, R7, Rs and Rg are each, independently, —O—or a—O—(C₂-C₆ alkylene)-SO₃ group, wherein at least one of R₁ to R₉ isindependently a-0-(C₂-C₆ alkylene)-SO,-group, preferably a —O—(CH₂)_(m)SO₃-group, wherein m is 2 to 6, preferably 2 to 4, (e.g.—OCH₂CH₂CH₂SO₃ —Or—OCH₂CH₂CH₂CH₂SO₃-); and S₁, S₂, S₃, S₄, S₅, S₆, S₇,S₈ and S₉ are each, independently, a pharmaceutically acceptable cationwhich includes, for example, H⁺, alkali metals (e.g. Li⁺, Na⁺, K⁺),alkaline earth metals (e.g., Ca⁺², Mg⁺²), ammonium ions and aminecations such as the cations of (C₁-C₆)-alkylamines, piperidine,pyrazine, (C₁-C₆)-alkanolamine and (C₄-C₈)-cycloalkanolamine.

Particularly suitable SAE-CD derivatives include those wherein n is 5 or6.

The SAE-CD used is available from CyDex Pharmaceuticals, Inc. (Lenexa,KS), and it is described in U.S. Pat. No. 5,376,645 and No. 5,134,127 toStella et al, the entire disclosures of which are hereby incorporated byreference. U.S. Pat. No. 3,426,011 to Parmerter et al. discloses anioniccyclodextrin derivatives having sulfoalkyl ether substituents. Lammerset al. (Reel. Trav. Chim. Pays-Bas (1972), 91(6), 733-742); (Staerke(1971), 23(5), 167-171) and Qu et al. (J. Inclusion Phenom. Macro.Chem., (2002), 43, 213-221) disclose sulfoalkyl ether derivatizedcyclodextrins. U.S. Pat. No. 6,153,746 to Shah et al. discloses aprocess for the preparation of sulfoalkyl ether cyclodextrinderivatives. An SAE-CD can be made according to the disclosures ofStella et al., Parmerter et al., Lammers et al. or Qu et al., and ifprocessed to remove the major portion (>50%) of the underivatized parentcyclodextrin, used according to the present invention. The SAE-CD cancontain from 0% to less than 50% wt. of underivatized parentcyclodextrin.

The terms “alkylene” and “alkyl,” as used herein (e.g., in the—O—(C₂-C₆-alkylene) SO₃— group or in the alkylamines), include linear,cyclic, and branched, saturated and unsaturated (i.e., containing onedouble bond) divalent alkylene groups and monovalent alkyl groups,respectively. The term “alkanol” in this text likewise includes bothlinear, cyclic and branched, saturated and unsaturated alkyl componentsof the alkanol groups, in which the hydroxyl groups may be situated atany position on the alkyl moiety. The term “cycloalkanol” includesunsubstituted or substituted (e.g., by methyl or ethyl) cyclic alcohols.

Exemplary embodiments of the SAE-CD derivative of the invention includederivatives of the Formula II (SAEx-α-CD), wherein “x” ranges from 1 to18; of the Formula III (SAEy-β-CD), wherein “y” ranges from 1 to 21; andof the Formula IV (SAEz-γ-CD), wherein “z” ranges from 1 to 24 such as:

SAEx-α-CD SAEy-β-CD SAEz-γ-CD Name SEEx-α-CD SEEy-β-CD SEEz-γ-CDSulfoethyl ether CD SPEx-α-CD SPEy-β-CD SPEz-γ-CD Sulfopropyl ether CDSBEx-α-CD SBEy-β-CD SBEz-γ-CD Sulfobutyl ether CD SPtEx-α-CD SPtEy-β-CDSPtEz-γ-CD Sulfopentyl ether CD SHEx-α-CD SHEy-β-CD SHEz-γ-CD Sulfohexylether CD

“SAE” represents a sulfoalkyl ether substituent bound to a cyclodextrin.The values “x”, “y” and “z” represent the average degree of substitutionas defined herein in terms of the number of sulfoalkyl ether groups perCD molecule.

Other exemplary SAE-CD derivatives include those of the formulaSAEx-R-CD (Formula 2), wherein SAE is sulfomethyl ether (SME),sulfoethyl ether (SEE), sulfopropyl ether (SPE), sulfobutyl ether (SBE),sulfopentyl ether (SPtE), or sulfohexyl ether (SHE); x (average orspecific degree of substitution) is 1-18, 1-21, 1-24, when R (ringstructure of parent cyclodextrin) is α, or γ, respectively, and CD iscyclodextrin.

Exemplary SAE-CD derivatives include SBE4-β-CD, SBE5.5-β-CD (Advasep®cyclodextrin), SBE7β-CD (CAPTISOL® cyclodextrin), SPE5.6-β-CD,SBE6.1-γ-CD and SBE7.6-γ-CD. Particularly suitable SAE-CD derivativesinclude SAE-β-CD and SAE-γ-CD.

The present invention provides compositions containing a mixture ofcyclodextrin derivatives, having the structure set out in the formulasabove, where the composition overall contains on the average at least 1and up to 3n+6 alkylsulfonic acid moieties per cyclodextrin molecule.The present invention also provides compositions containing a singletype of cyclodextrin derivative, or at least 50% of a single type ofcyclodextrin derivative.

It should be understood that other SAE-CD compounds of the aboveformulas may be used in the liquid formulation of the invention. Theseother SAE-CD formulations differ from SBE7-β-CD in their degree ofsubstitution by sulfoalkyl groups, the number of carbons in thesulfoalkyl chain, their molecular weight, the number of glucopyranoseunits contained in the base cyclodextrin used to form the SAE-CD and ortheir substitution patterns. In addition, the derivatization ofcyclodextrin with sulfoalkyl groups occurs in a controlled, although notexact manner. For this reason, the degree of substitution is actually anumber representing the average number of sulfoalkyl groups percyclodextrin (for example, SBE7-β-CD, has an average of 7 substitutionsper cyclodextrin). In addition, the regiochemistry of substitution ofthe hydroxyl groups of the cyclodextrin is variable with regard to thesubstitution of specific hydroxyl groups of the hexose ring. For thisreason, sulfoalkyl substitution of the different hydroxyl groups islikely to occur during manufacture of the SAE-CD, and a particularSAE-CD will possess a preferential, although not exclusive or specific,substitution pattern. Given the above, the molecular weight of aparticular SAE-CD may vary from batch to batch and will vary from SAE-CDto SAE-CD. All of these variations can lead to changes in thecomplexation equilibrium constant which in turn will affect the requiredmolar ratios of the SAE-CD to clopidogrel. The equilibrium constant isalso somewhat variable with temperature and allowances in the ratio arerequired such that the agent remains solubilized during the temperaturefluctuations that can occur during manufacture, storage, transport, anduse. The equilibrium constant is also variable with pH and allowances inthe ratio are required such that the agent remains solubilized during pHfluctuations that can occur during manufacture, storage, transport, anduse. The equilibrium constant is also variable by the presence of otherexcipients (e.g., buffers, preservatives, antioxidants). Accordingly,the ratio of SAE-CD/clopidogrel may need to be varied (±) from theratios set forth herein in order to compensate for the above-mentionedvariables.

In some embodiments, the cyclodextrin derivatives of the presentinvention can be obtained as purified compositions, i.e., compositionscontaining at least 90 wt. % or 95 wt. % of cyclodextrin derivative(s)in terms of the total amount of cyclodextrin present, the balance ofcyclodextrin comprising unreacted parent cyclodextrin. In a preferredembodiment, purified compositions containing at least 98 wt. %cyclodextrin derivative(s) are obtained. In some of the compositions ofthe invention unreacted cyclodextrin has been substantially removed,with the remaining impurities (i.e., <5 wt. % of composition) beinginconsequential to the performance of the cyclodextrinderivative-containing composition.

According to other embodiments, the amount of unreacted parentcyclodextrin present in the SAE-CD is up to about or less than about 50%wt. of the SAE-CD, less than about 40% wt., less than 30% wt., or lessthan 20% wt. based upon the total dry weight of cyclodextrin.

By “clopidogrel/SAE-CD complex” is generally meant a clathrate orinclusion complex of a sulfoalkyl ether cyclodextrin derivative of theformula (1) and clopidogrel. The complex can be a binary or ternarycomplex (the salt form of clopidogrel is complexed). The ratio ofSAE-CD:clopidogrel present in the molecular complex is in the range ofabout 1:1, on a molar basis. However, it should be understood that themolar ratio of SAE-CD to clopidogrel in the solution, as a whole, willbe higher such that the SAE-CD will generally be, but need not be,present in molar excess over the clopidogrel. The amount of excess willbe determined by the intrinsic solubility of the clopidogrel form, theexpected dose of the clopidogrel form, and the binding constant forinclusion complexation between the specific clopidogrel form and thespecific SAE-CD.

By “major portion” is meant at least about 50% by weight of thetherapeutic compound. In various specific embodiments, greater than 50%,60%, 75%, 90% or 95% by weight of the clopidogrel can be complexed withan SAE-CD while in the pharmaceutical formulation. The actual percent ofdrug that is complexed will vary according to the complexationequilibrium constant characterizing the complexation of a specificSAE-CD to clopidogrel and to the concentrations of SAE-CD andclopidogrel available for complexation.

FIG. 1 depicts the results of a phase solubility study per the method ofHiguchi et al. in Phase Solubility Techniques, in Advances in AnalyticalChemistry and Instrumentation (Ed. CN. Reilly, John Wiley & Sons Inc.,Vol. 4 (1965), pg. 117-212) comparing the dissolution power of variouscyclodextrins (SBE6.6-β-CD, SBE5.5-β-CD SBE7.6-γ-CD, SPE5.8-CC-CD,SPE5.6-γ-CD, SPE5.4-γ-CD, SBE3.9-CC-CD, SPE5.0-β-CD, and SBE2.4-CC-CD)as compared to HP-β-CD and α-CD for binding with clopidogrel on a molarbasis. The study was conducted in a titration apparatus that maintainedthe pH of the solutions at −5.5. Samples were analyzed for clopidogrelcontent by HPLC with UV detection.

The results detailed in FIG. 1 indicate that SAE-CD generallyoutperforms HP-β-CD. In particular, SBE7-β-CD provides the highestdegree of dissolution under the conditions tested. The SAE-β-CD andSAE-γ-CD derivatives are particularly useful for solubilizingclopidogrel. It should be noted that the performance of the variousSAE-CD derivatives can be improved by varying test conditions and/orsolution properties.

The thermal and hydrolytic stability of clopidogrel in aqueous liquidformulations according to the invention was evaluated at varioustemperatures and pH's and phosphate buffer concentrations as detailed inExample 19. A similar evaluation was conducted for liquid formulationscontaining HP-β-CD rather than SAE-CD. The studies utilized equimolaramounts of the cyclodextrin derivatives, even though substantially moreclopidogrel could be solubilized by the SAE-CD than with the HP-β-CD.These results (FIGS. 2 a-2 b) indicate a slower rate of thermal orhydrolytic degradation of clopidogrel in the presence of SAE-CD ascompared to degradation of clopidogrel in the presence of HP-β-CD.

Another measure of the stability of clopidogrel is its rate of chiralinversion when present in a solution. The stability of clopidogrel in anaqueous liquid formulation according to the invention was evaluated atvarious temperatures and pH's and phosphate buffer concentrations asdetailed in Example 19. A similar evaluation was conducted for liquidformulations containing HP-β-CD rather than SAE-CD. The rate of chiralconversion from (S)-clopidogrel to (R)-clopidogrel is dependent upon pHof the medium as is seen by comparing the ratio of enantiomers (R:S) inthe pH˜5.5 and the pH˜8 formulations (FIGS. 3 a-3 b). It should be notedthat the formulations with SAE-CD demonstrate a significant improvementin clopidogrel stabilization over those containing HP-β-CD, i.e.substantially less conversion of S-clopidogrel to R-clopidogrel. Theresults (FIGS. 3 a-3 b) establish the superiority of SAE-CD over HP-β-CDin stabilizing clopidogrel against chiral inversion in solution.

The photochemical stability of two SAE-CD based formulations and anHP-β-CD based formulation was evaluated as detailed in Example 19. Aportion of each formulation was exposed to fluorescent light over aperiod of nine days. At various time points, aliquots of solution werewithdrawn and analyzed by HPLC to determine their impurity profile andisomeric content.

FIGS. 4 a and 4 b depict the results of a stability assay to determinethe impact of the fluorescence irradiation upon the chiral inversion ofthe clopidogrel. The results establish the unexpectedly greaterstabilization of clopidogrel by SBE-β-CD toward chiral inversion (andultimate racemization) as compared to the limited degree ofstabilization provided by HP-β-CD.

The chemical stability of the liquid formulations of the invention, interms of formation of a precipitate, can be enhanced by adjusting the pHof the liquid carrier. The chemical stability can also be enhanced byconverting the liquid formulation to a solid or powder formulation.

The pH of water (for distilled and/or deionized water) in which acomposition or formulation of the invention is placed for reconstitutioncan generally range from 1 to 8, provided the water does not include asubstantial amount of a buffer or excludes a buffer; however,compositions or formulations having higher or lower pH values can alsobe prepared. The pH can be varied according to the intended mode ofadministration to a subject. The pH of a parenteral formulation cangenerally range from about pH 4 to about pH 8 or about pH 4 to about pH6. The pH of an oral formulation will generally range from about pH 1 toabout pH 8, about pH 4 to about pH 8, about pH 4 to about pH 6, about pH1 to about pH 3, or about pH 1 to about pH 4. The pH solubility profile(FIG. 5) indicates that the solubility of clopidogrel (in the absence ofa cyclodextrin derivative) is dependent upon pH. At a solution pH ofabout 3, the solubility of clopidogrel bisulfate is about 5.0 mg/ml.Below about pH 3, the solubility of clopidogrel increases dramatically,and above about pH 3, the solubility of clopidogrel decreases asfollows:

Solubility of Clopidogrel Solution pH (mg/ml; approximate) 3.5 1.79 4.00.279 4.5 0.171 5.12 0.083 6.60 0.082

The phase solubility profiles of clopidogrel in SAE-CD at pH˜5.5 (FIGS.6 a and 6 b) indicate that the solubility of the clopidogrel isdependent upon the cyclodextrin content. The slope of the line in FIG. 6b is about 0.15 indicating that the approximate minimum molar ratio ofSBE-β-CD to clopidogrel required to dissolve the clopidogrel at pH 5.5is at least about 6:1. As the concentration of SAE-CD is increased,greater amounts of clopidogrel can be dissolved and more concentratedsolutions of clopidogrel can be prepared. Thus, concentratedformulations of clopidogrel can be prepared by using higherconcentrations of SAE-CD. For parenteral applications, concentratedsolutions would reduce the administration volume and potentially allowfor faster administration. Concentrated solutions would also allow forperoral administration of doses in smaller volumes, and the requiredhigher SAE-CD concentrations would provide improved antimicrobial actionas well as potentially improved taste masking. In addition, dilutesolutions can also be prepared, and the amount of cyclodextrin requiredwould be reduced. The dilute solutions would allow for sloweradministration of the formulation and potentially better control of thedose administered over time.

The invention also provides a taste-masked oral formulation comprisingsulfoalkyl ether cyclodextrin, clopidogrel, a pharmaceuticallyacceptable carrier and optionally other ingredients. In someembodiments, a taste-masked formulation comprises SAE-CD, clopidogrel, apharmaceutically acceptable carrier, and aqueous solvent, suspendingagent, buffer, surfactant, cosolvent and/or a flavoring agent such asmannitol, glucose, sucrose, xylitol and others known to those ofordinary skill in the art. Examples 15 and 17 describe exemplarytaste-masked formulations comprising clopidogrel bisulfate, SAE-CD and asugar, such as mannitol or D-glucose, respectively. The SAE-CD andclopidogrel molar ratio required may depend upon its mode ofadministration, the pH of a formulation containing the same, the pH ofan intended environment of delivery. Typically the molar ratio can bewithin the range of 6:1 and 8:1, 6:1 to 10:1, 5:1 to 12:1, 4:1 to 15:1,5:1 to 14:1, 6:1 to 13:1, or 6:1 to 12.5:1; and/or at least 0.05:1, atleast 0.25:1, at least 0.2:1, at least 0.5:1, at least 1:1, at least2:1, at least 3:1, at least 4:1, at least 5:1, at least 6:1, at least7:1, or at least 8:1; and/or at most 100:1, at most 75:1, at most 50:1,at most 40:1, at most 35:1, at most 30:1, at most 25:1, at most 20:1, atmost 15:1, at most 14:1 at most 12.5:1, at most 12:1, at most 10:1, atmost 8:1, at most 6:1, at most 5:1, at most 4:1, at most 3:1, or at most2:1. The reconstituted pH of the formulations can be low, e.g. betweenpH 1-2, which is suitable for oral use. The formulation of the inventioncan comprise clopidogrel in combination with a second therapeutic agent.The second therapeutic agent can be selected from the group consistingof: nonsteroidal antiinflamatory drugs like piroxicam or aspirin;anticoagulants (e.g. warfarin, antithrombin III, unfractionated heparin,heparinoids like danaparoid, low molecular weight heparins likeenoxaparin, selective factor Xa inhibitors like fondaparinux, and directthrombin inhibitors like argatroban and bivalirudin); antiplateletagents (e.g. anagrelide, dipyridamole, aggregation inhibitors likecilostazol and cangrelor, and glycoprotein Ilb/IIIa inhibitors likeeptifibatide and tirofiban); antisickling agents like hydroxyurea;hemorrheologic agents like pentoxifylline; and thrombolytic agents (e.g.biologic response modifiers like drotrecogin alpha and pexelizumab,thrombolytic enzymes like streptokinase and urokinase, and tissueplasminogen activators like alteplase and tenecteplase). This isdemonstrated in Examples 24 and 25, where clopidogrel is formulated incombination with tirofiban hydrochloride monohydrate and enoxaprinsodium.

In some embodiments, the second therapeutic agent is used to treatsickle cell disease. Exemplary of such second therapeutic agents are: 1)drugs included in management protocols for sickle cell patientsexperiencing pain, fever-febrile illness acute chest syndrome, acutesplenic sequestration, aplastic crisis, acute stroke or neurologicevents; 2) folic acid supplementation, e.g. as part of a hydroxyureatreatment protocol; 3) NSAID, such as ibuprofen, acetaminophen, aspirin,codeine, morphine, hydromorphone, and ketorolac; 4) antibiotic, such aspenicillin, penicillin derivatives, cephalosporins (such as ceftriaxone,cefotaxime, and others known to those in the art), macrolides (suchazithromycin, erythromycin), clindamycin and vancomycin; 5)deferoximine, such as part of an iron chelation treatment protocol dueto chronic transfusion requirements; 6) bronchodilators; 7) diuretic,such as furosemide; 8) anxiolytics, such as lorazepam, midazolam orhydroxyzine pamoate; 9) α-agonist, such as etilefrine, phenylephrine,epinephrine, phenylpropanolamine, pseudoephedrine, or terbutaline; 10)hydralazine; 11) pentoxifylline; 12) diltiazem; 13) gonatropin-releasinghormone analog such as leuprolide or flutamide; 14) diethylstilbestrol;and 15) combinations thereof.

The amount of the other drug present and thus the ratio of clopidogrelto the other drug present will depend upon the desired clinical effect.However, guidance as to the relative doses of each drug can be obtainedfrom the resources of regulatory agencies such as the U.S. Food and DrugAdministration, or other similarly recognized authority in Canada(Health Canada), Mexico (Mexico Department of Health), Europe (EuropeanMedicines Agency (EMEA)), South America (in particular in Argentina(Administración Nacional de Medic amentos, Alimentos y Tecnologia Medica(ANMAT)) and Brazil (Ministerio da Sa{acute over (υ)}de)), Australia(Department of Health and Ageing), Africa (in particular in South Africa(Department of Health) and Zimbabwe (Ministry of Health and ChildWelfare)), or Asia (in particular Japan (Ministry of Health, Labour andWelfare), Taiwan (Executive Yuans Department of Health), and China(Ministry of Health People's Republic of China)).

When clopidogrel is present with another drug in an aqueous solutioncontaining SAE-CD, there is a potential for competitive binding of theclopidogrel and other drug with SAE-CD. The extent to which competitivebinding might occur can be predicted to some degree by comparison of thebinding constant of clopidogrel versus that of the other drug. Thegreater a binding constant of a drug for a particular cyclodextrin, themore tightly bound the drug is to the cyclodextrin and the greater theamount of drug that can be solubilized by the cyclodextrin derivative.If the other drug has a much greater binding constant for SAE-CD thandoes clopidogrel, then the other drug will likely competitively bindSAE-CD. If the other drug has a much smaller binding constant for SAE-CDthan does clopidogrel, then the other drug will likely not competitivelybind SAE-CD.

If the binding constant of the other drug for SAE-CD approximates thatof clopidogrel for SAE-CD, then the extent to which competitive bindingoccurs will be driven more by the relative molar ratio of the two drugsthan by the binding constant of the two drugs. In other words, if theother drug is administered at a substantially higher molar concentrationthan is clopidogrel, then it will more competitively bind SAE-CD than ifthe other drug was administered at a substantially lower molarconcentration than clopidogrel.

Exemplary formulations comprising SAE-CD, clopidogrel bisulfate, aqueousbuffer, and aspirin were prepared according to Example 22. In thisexample, excess clopidogrel bisulfate was added to solutions containingfixed amounts of SAE-CD and varying amounts of aspirin. FIG. 7 depictsthe results of a competitive binding study. The results indicate thataspirin binds competitively with SAE-CD when placed in a solutioncontaining SAE-CD and clopidogrel salt. Thus, in a formulationcontaining both clopidogrel and aspirin, the formulation may or may notrequire additional SAE-CD to solubilize both ingredients than would berequired to solubilize either component individually.

Although not necessary, the formulation of the present invention mayinclude a antioxidant, acidifying agent, alkalizing agent, bufferingagent, bulking agent, cryoprotectant, density modifier, electrolyte,flavors, fragrance, glucose, stabilizer, plasticizer,solubility-enhancing agent, sweeteners, surface tension modifier,volatility modifier, viscosity modifier, other excipients known by thoseof ordinary skill in the art for use in pharmaceutical formulations, ora combination thereof.

A complexation-enhancing agent can be added to the aqueous liquidformulation of the invention. A complexation-enhancing agent is acompound, or compounds, that enhance(s) the complexation of clopidogrelwith the SAE-CD. When the complexation-enhancing agent is present, therequired ratio of SAE-CD to clopidogrel may need to be changed such thatless SAE-CD is required. Suitable complexation enhancing agents includeone or more pharmacologically inert water soluble polymers, hydroxyacids, and other organic compounds typically used in liquid formulationsto enhance the complexation of a particular agent with cyclodextrins.Suitable water soluble polymers include water soluble natural polymers,water soluble semisynthetic polymers (such as the water solublederivatives of cellulose) and water soluble synthetic polymers. Thenatural polymers include polysaccharides such as inulin, pectins, alginderivatives and agar, and polypeptides such as casein and gelatin. Thesemi-synthetic polymers include cellulose derivatives such asmethylcellulose, hydroxyethyl cellulose, hydroxypropyl cellulose, theirmixed ethers such as hydroxypropyl methylcellulose and other mixedethers such as hydroxyethyl ethylcellulose, hydroxypropylethylcellulose, hydroxypropyl methylcellulose phthalate andcarboxymethylcellulose and its salts, especially sodiumcarboxymethylcellulose. The synthetic polymers include polyoxyethylenederivatives (polyethylene glycols) and polyvinyl derivatives (polyvinylalcohol, polyvinylpyrrolidone and polystyrene sulfonate) and variouscopolymers of acrylic acid (e.g. carbomer). Suitable hydroxy acidsinclude by way of example, and without limitation, citric acid, malicacid, lactic acid, and tartaric acid and others known to those ofordinary skill in the art.

Hydrophilic polymers can be used to improve the performance offormulations containing a cyclodextrin. Loftsson has disclosed a numberof polymers suitable for combined use with a cyclodextrin (underivatizedor derivatized) to enhance the performance and/or properties of thecyclodextrin. Suitable polymers are disclosed in Pharmazie (2001),56(9), 746-747; International Journal of Pharmaceutics (2001), 212(1),29-40; Cyclodextrin: From Basic Research to Market, InternationalCyclodextrin Symposium, 10th, Ann Arbor, Mich., United States, May21-24, 2000 (2000), 10-15 (Wacker Biochem Corp.: Adrian, Mich.); PCTInternational Publication No. WO 9942111; Pharmazie, (1998), 53(11),733-740; Pharm. Technol. Eur, (1997), 9(5), 26-34; 1 J. Pharm. Sci(1996), 85(10), 1017-1025; European Patent Application EP0579435;Proceedings of the International Symposium on Cyclodextrins, 9th,Santiago de Comostela, Spain, May 31-Jun. 3, 1998 (1999), 261-264(Editor(s): Labandeira, J. J. Torres; Vila-Jato, J. L. Kluwer AcademicPublishers, Dordrecht, Neth); S. T. P. Pharma Sciences (1999), 9(3),237-242; ACS Symposium Series (1999), 737 (Polysaccharide Applications),24-45; Pharmaceutical Research (1998), 15(11), 1696-1701; DrugDevelopment and Industrial Pharmacy (1998), 24(4), 365-370;International Journal of Pharmaceutics (1998), 163(1-2), 115-121; Bookof Abstracts, 216th ACS National Meeting, Boston, Aug. 23-27 (1998),CELL-016, American Chemical Society; Journal of Controlled Release,(1997), 44/1 (95-99); Pharm. Res. (1997), 14(11), 5203; InvestigativeOphthalmology & Visual Science, (1996), 37(6), 1199-1203; Proceedings ofthe International Symposium on Controlled Release of Bioactive Materials(1996), 23rd, 453-454; Drug Development and Industrial Pharmacy (1996),22(5), 401-405; Proceedings of the International Symposium onCyclodextrins, 8th, Budapest, Mar. 31-Apr. 2, (1996), 373-376.(Editor(s): Szejtli, J.; Szente, L. Kluwer: Dordrecht, Neth.);Pharmaceutical Sciences (1996), 2(6), 277-279; European Journal ofPharmaceutical Sciences, (1996) 4(SUPPL.), S144; Third European Congressof Pharmaceutical Sciences Edinburgh, Scotland, UK Sep. 15-17, 1996;Pharmazie, (1996), 51(1), 39-42; Eur. J Pharm. Sci (1996), 4 (Suppl.),S143; U.S. Pat. No. 5,472,954 and No. 5,324,718; International Journalof Pharmaceutics (Netherlands), (Dec. 29, 1995) 126, 73-78; Abstracts ofPapers of the American Chemical Society, (02 Apr. 1995) 209(1), 33-CELL;European Journal of Pharmaceutical Sciences, (1994) 2, 297-301;Pharmaceutical Research (New York), (1994) 11(10), S225; InternationalJournal of Pharmaceutics (Netherlands), (Apr. 11, 1994) 104, 181-184;and International Journal of Pharmaceutics (1994), 110(2), 169-77, theentire disclosures of which are hereby incorporated by reference.

Other suitable polymers are well-known excipients commonly used in thefield of pharmaceutical formulations and are included in, for example,Remington's Pharmaceutical Sciences, 18th Edition, Alfonso R. Gennaro(editor), Mack Publishing Company, Easton, Pa., 1990, pp. 291-294;Alfred Martin, James Swarbrick and Arthur Commarata, Physical Pharmacy.Physical Chemical Principles in Pharmaceutical Sciences, 3rd edition(Lea & Febinger, Philadelphia, Pa., 1983, pp. 592-638); A. T. Florenceand D. Altwood, (Physicochemical Principles of Pharmacy, 2nd Edition,MacMillan Press, London, 1988, pp. 281-334. The entire disclosures ofthe references cited herein are hereby incorporated by references. Stillother suitable polymers include water-soluble natural polymers,water-soluble semi-synthetic polymers (such as the water-solublederivatives of cellulose) and water-soluble synthetic polymers. Thenatural polymers include polysaccharides such as inulin, pectin, alginderivatives (e.g. sodium alginate) and agar, and polypeptides such ascasein and gelatin. The semi-synthetic polymers include cellulosederivatives such as methylcellulose, hydroxyethylcellulose,hydroxypropyl cellulose, their mixed ethers such as hydroxypropylmethylcellulose and other mixed ethers such as hydroxyethylethylcellulose and hydroxypropyl ethylcellulose, hydroxypropylmethylcellulose phthalate and carboxymethylcellulose and its salts,especially sodium carboxymethylcellulose. The synthetic polymers includepolyoxyethylene derivatives (polyethylene glycols) and polyvinylderivatives (polyvinyl alcohol, polyvinylpyrrolidone and polystyrenesulfonate) and various copolymers of acrylic acid (e.g. carbomer). Othernatural, semi-synthetic and synthetic polymers not named here which meetthe criteria of water solubility, pharmaceutical acceptability andpharmacological inactivity are likewise considered to be within theambit of the present invention.

A solubility-enhancing agent can be added to the aqueous liquidformulation of the invention. A solubility-enhancing agent is acompound, or compounds, that enhance(s) the solubility of clopidogrel inthe liquid formulation. When a complexation-enhancing agent is present,the ratio of SAE-CD to clopidogrel may need to be changed such that lessSAE-CD is required. Suitable solubility enhancing agents include one ormore organic solvents, detergents, soaps, surfactants and other organiccompounds typically used in oral solution formulations to enhance thesolubility of a particular agent. Suitable organic solvents include, forexample, ethanol, glycerin, polyethylene glycols, propylene glycol,poloxomers, and others known to those of ordinary skill in the art.

Surfactants, which are considered solubility-enhancing agents, can beused to enhance solubilization of clopidogrel and reduce the amount ofSAE-CD required to dissolve the clopidogrel. Exemplary formulationscomprising SAE-CD, clopidogrel bisulfate, aqueous buffer, andsurfactants were prepared according to Example 14. There was a markedincrease in solubility of clopidogrel when using both polyethyleneglycol-15-hydroxy stearate (Solutol®) and polysorbate 80 (Tween 80®).Other surfactants such as and not limited to; Cremophor®, polysorbate20, 40, 60 and 80, Solutol®, Labrasol®, poloxamers, polyethylene glycolderivatives, cholate acids and their derivatives can be used and othersknown to individuals of ordinary skill in the art can also be used incombination with the SAE-CD. Exemplary solubility enhancing agents aredisclosed in U.S. Pat. No. 6,451,339; however, other solubilityenhancing agents used in the pharmaceutical industry can be used in theformulation of the invention.

The combined use of SAE-CD and surfactant may result in either asynergistic, additive or a negative effect on the solubility of theclopidogrel. The observed effect can be dependent upon the concentrationof surfactant, in that using the same surfactant at differentconcentrations can produce different effects as is demonstrated inExample 14.

As shown in Tables 14a and 14b, Tween 80 shows an additive effect whenused at concentrations of 1-2% (w/v) but a negative effect when used at0.1, 0.5, 5 and 10% (w/v). Solutol (Tables 14c and 14d) shows asynergistic effect in the solubility at concentrations of between 1-5%(w/v). There is no change in the solubility at a concentration of 10%(w/v) of Solutol with and without CAPTISOL. Without being held bound toa particular mechanism, the negative solubility observed whensurfactants are used with cyclodextrins may be due to the surfactantmolecule acting as a competitive inhibitor in the solubilization of thedrug by the complexant, in this case cyclodextrin. Similarly thecomplexant (the cyclodextrin) “pulls” the surfactant out of solution,making it unavailable for solubilizing the drug.

As used herein, the term “flavor” is intended to mean a compound used toimpart a pleasant flavor and often odor to a pharmaceutical preparation.Exemplary flavoring agents or flavorants include synthetic flavor oilsand flavoring aromatics and/or natural oils, extracts from plants,leaves, flowers, fruits and so forth and combinations thereof. These mayalso include cinnamon oil, oil of wintergreen, peppermint oils, cloveoil, bay oil, anise oil, eucalyptus, thyme oil, cedar leave oil, oil ofnutmeg, oil of sage, oil of bitter almonds and cassia oil. Other usefulflavors include vanilla, citrus oil, including lemon, orange, grape,lime and grapefruit, and fruit essences, including apple, pear, peach,strawberry, raspberry, cherry, plum, pineapple, apricot and so forth.Flavors which have been found to be particularly useful includecommercially available strawberry, orange, grape, cherry, vanilla, mintand citrus flavors and mixtures thereof. The amount of flavoring maydepend on a number of factors, including the organoleptic effectdesired. Flavors will be present in any amount as desired by those ofordinary skill in the art. Particularly flavors are the strawberry andcherry flavors and citrus flavors such as orange.

As used herein, the term “sweetener” is intended to mean a compound usedto impart sweetness to a preparation. Such compounds include, by way ofexample and without limitation, aspartame, dextrose, glycerin, mannitol,saccharin sodium, sorbitol, xylitol, fructose, high fructose corn syrup,maltodextrin, sucralose, sucrose, other materials known to one ofordinary skill in the art, and combinations thereof.

As used herein, a fragrance is a relatively volatile substance orcombination of substances that produces a detectable aroma, odor orscent. Exemplary fragrances include those generally accepted as FD&C.

As used herein, the term “alkalizing agent” is intended to mean acompound used to provide alkaline medium. Such compounds include, by wayof example and without limitation, ammonia solution, ammonium carbonate,diethanolamine, monoethanolamine, potassium hydroxide, sodium borate,sodium carbonate, sodium bicarbonate, sodium hydroxide, triethanolamine,diethanolamine, organic amine base, alkaline amino acids and trolamineand others known to those of ordinary skill in the art.

As used herein, the term “acidifying agent” is intended to mean acompound used to provide an acidic medium. Such compounds include, byway of example and without limitation, acetic acid, acidic amino acids,citric acid, fumaric acid and other alpha hydroxy acids, hydrochloricacid, ascorbic acid, phosphoric acid, sulfuric acid, tartaric acid andnitric acid and others known to those of ordinary skill in the art.

As used herein, the term “preservative” is intended to mean a compoundused to prevent the growth of microorganisms. Such compounds include, byway of example and without limitation, benzalkonium chloride,benzethonium chloride, benzoic acid, benzyl alcohol, cetylpyridiniumchloride, chlorobutanol, phenol, phenylethyl alcohol, phenylmercuricnitrate, phenylmercuric acetate, thimerosal, metacresol, myristylgammapicolinium chloride, potassium benzoate, potassium sorbate, sodiumbenzoate, sodium propionate, sorbic acid, thymol, and methyl, ethyl,propyl, or butyl parabens and others known to those of ordinary skill inthe art.

As used herein, the term “antioxidant” is intended to mean an agentwhich inhibits oxidation and thus is used to prevent the deteriorationof preparations by the oxidative process. Such compounds include by wayof example and without limitation, acetone, sodium bisulfate, ascorbicacid, ascorbyl palmitate, citric acid, butylated hydroxyanisole,butylated hydroxytoluene, hydrophosphorous acid, monothioglycerol,propyl gallate, sodium ascorbate, sodium citrate, sodium sulfide, sodiumsulfite, sodium bisulfite, sodium formaldehyde sulfoxylate, thioglycolicacid, sodium metabisulfite, EDTA (edetate), pentetate and others knownto those of ordinary skill in the art.

As used herein, the term “buffering agent” is intended to mean acompound used to resist change in pH upon dilution or addition of acidor alkali. Such compounds include, by way of example and withoutlimitation, acetic acid, sodium acetate, adipic acid, benzoic acid,sodium benzoate, citric acid, maleic acid, monobasic sodium phosphate,dibasic sodium phosphate, lactic acid, tartaric acid, glycine, potassiummetaphosphate, potassium phosphate, monobasic sodium acetate, sodiumbicarbonate, sodium tartrate and sodium citrate anhydrous and dihydrateand others known to those of ordinary skill in the art.

As used herein, the term “stabilizer” is intended to mean a compoundused to stabilize a therapeutic agent against physical, chemical, orbiochemical process that would otherwise reduce the therapeutic activityof the agent. Suitable stabilizers include, by way of example andwithout limitation, albumin, sialic acid, creatinine, glycine and otheramino acids, niacinamide, sodium acetyltryptophonate, zinc oxide,sucrose, glucose, lactose, sorbitol, mannitol, glycerol, polyethyleneglycols, sodium caprylate and sodium saccharin and others known to thoseof ordinary skill in the art.

As used herein, the term “viscosity modifier” is intended to mean acompound or combination of compounds capable of increasing or decreasingthe viscosity of the liquid formulation. Some of the polymers disclosedherein can be used as viscosity modifiers.

As used herein, the term “tonicity modifier” is intended to mean acompound or compounds that can be used to adjust the tonicity of theliquid formulation. Suitable tonicity modifiers include glycerin,lactose, mannitol, dextrose, sodium chloride, sodium sulfate, sorbitol,trehalose and others known to those or ordinary skill in the art.

As used herein, the term “antifoaming agent” is intended to mean acompound or compounds that prevents or reduces the amount of foamingthat forms on the surface of the liquid formulation. Suitableantifoaming agents include by way of example and without limitation,dimethicone, simethicone, octoxynol and others known to those ofordinary skill in the art.

As used herein, the term “bulking agent” is intended to mean a compoundused to add bulk to the reconstitutable solid and/or assist in thecontrol of the properties of the formulation during preparation. Suchcompounds include, by way of example and without limitation, dextran,trehalose, sucrose, polyvinylpyrrolidone, lactose, inositol, mannitol,sorbitol, dimethylsulfoxide, glycerol, albumin, calcium lactobionate,and others known to those of ordinary skill in the art.

As used herein, the term “cryoprotectant” is intended to mean a compoundused to protect an active therapeutic agent from physical or chemicaldegradation during lyophilization. Such compounds include, by way ofexample and without limitation, dimethyl sulfoxide, glycerol, trehalose,propylene glycol, polyethylene glycol, and others known to those ofordinary skill in the art.

It should be understood, that compounds used in the pharmaceutical artsgenerally serve a variety of functions or purposes. Thus, if a compoundnamed herein is mentioned only once or is used to define more than oneterm herein, its purpose or function should not be construed as beinglimited solely to that named purpose(s) or function(s).

The liquid formulation of the invention can be prepared by numerousdifferent methods. According to one method, a first aqueous solutioncomprising SAE-CD is prepared. Then, a second solution comprisingclopidogrel is prepared. Finally, the first and second solutions aremixed to form the liquid formulation. The first and second solutions canindependently comprise other excipients and agents described herein.Additionally, the second solution can be water and/or an organicsolvent-base solution. Another method of preparation is similar to theabove-described method except that the clopidogrel is added directly tothe first solution without the formation of a second solution. A thirdmethod of preparing the liquid formulation is similar to theabove-described first method except that the SAE-CD is added directly toan aqueous second solution containing the clopidogrel without formationof the first solution. A fourth method of preparing the liquidformulation comprises the steps of adding an aqueous solution comprisingclopidogrel to a powdered or particulate SAE-CD and mixing the solutionuntil the SAE-CD has dissolved. A fifth method of preparing the liquidformation comprises the steps of adding the clopidogrel directly to thepowdered or particulate SAE-CD and then adding an aqueous solution andmixing until the SAE-CD and clopidogrel has dissolved. A sixth methodfor preparing the liquid formation comprises the steps of heating eitherthe first solution or heating the second solution, or heating acombination thereof of any solutions described in the above methodsfollowed by the step of cooling the respectively heated solution. Aseventh method for preparing the liquid formation comprises the step ofadjusting the pH of either the first solution or adjusting the pH of thesecond solution or adjusting the pH of a combination of either solutionsdescribed in any of the above methods. An eighth method comprises thesteps of creating the liquid formulation by any of the above-describedmethods followed by the step of isolating a solid material bylyophilization, drum drying, spray-drying, spray freeze-drying,vacuum-drying, antisolvent precipitation or a process utilizing asupercritical or near supercritical fluid. Any of the above solutionscan contain other pharmaceutical excipients or ingredients as describedherein.

Some embodiments of the method of preparing the liquid formulationfurther comprise the step(s) of: 1) filtering the formulation through afiltration medium wherein the pore size is about 5 μm or smaller; 2)sterilizing the liquid formulation by irradiation; 3) sterilizing theliquid formulation by treatment with ethylene oxide; 4) isolating asterile powder from the sterilized liquid formulation; 5) purging theliquid with an inert gas to reduce the amount of dissolved oxygen in theliquid; and/or 6) one or more of the solutions used to prepare theliquid formulation is heated.

The invention provides pharmaceutical kits. In some embodiments, apharmaceutical kit comprises a first chamber containing a liquid vehicleand a second chamber containing a reconstitutable solid pharmaceuticalcomposition as described herein. The first and second chamber can beintegral or engaged or assembled to form a container with at least twochambers, or the chambers can be separate to form separate containers.The liquid vehicle comprises an aqueous liquid carrier such as water,dextrose, saline, lactated Ringer's solution, or any otherpharmaceutically acceptable aqueous liquid vehicles for the preparationof a liquid pharmaceutical compound. In some embodiments, the kitcomprises a container comprising a reconstitutable solid pharmaceuticalcomposition, and a container comprising a liquid carrier. In otherembodiments, the kit comprises a container comprising at least twochambers, wherein a chamber comprises a reconstitutable solidpharmaceutical composition, and another chamber comprises a liquidcarrier.

In some embodiments, the kit comprises a first chamber comprising apharmaceutical composition comprising an SAE-CD, and a second chambercomprising a pharmaceutical composition comprising clopidogrel. Thefirst and second chamber can be integral or engaged or assembled to forma container with at least two chambers, or the chambers can be separateto form separate containers. The first and second compositions can bemixed with a liquid carrier, which may or may not be included with thekit, and formulated as a liquid dosage form prior to administration to asubject. Either one or both of the first and second pharmaceuticalcompositions can comprise additional pharmaceutical excipients. The kitis available in various forms. In a first kit, the first and secondpharmaceutical compositions are provided in separate containers orseparate chambers of a container having two or more chambers. The firstand second pharmaceutical compositions may be independently provided ineither solid or powder or liquid form. For example, the SAE-CD can beprovided in a reconstitutable powder form and clopidogrel can beprovided in powdered form. According to one embodiment, the kit wouldfurther comprise a pharmaceutically acceptable liquid carrier used tosuspend and dissolve the first and/or second pharmaceuticalcompositions. Alternatively, a liquid carrier is independently includedwith the first and/or second pharmaceutical composition. The liquidcarrier, however, can also be provided in a container or chamberseparate from the first and second pharmaceutical compositions. Asabove, the first pharmaceutical composition, the second pharmaceuticalcomposition and the liquid carrier can independently comprise apreservative, an antioxidant, a buffering agent, an acidifying agent, anelectrolyte, another therapeutic agent, an alkalizing agent, anantimicrobial agent, an antifungal agent, a solubility enhancing agent,a viscosity modifying agent, a flavoring agent, a sweetening agent or acombination thereof.

Some embodiments of the kit include those wherein: 1) the first andsecond pharmaceutical compositions are contained in separate containersor separate chambers of a container having two or more chambers; 2) thekit further comprises a separate pharmaceutically acceptable liquidcarrier; 3) a liquid carrier is independently included upon eachoccurrence with the first and/or second pharmaceutical composition; 4)containers for the pharmaceutical compositions are independentlyselected at each occurrence from an evacuated container, bag, pouch,vial, bottle, or any pharmaceutically acceptable device known to thoseskilled in the art for the delivery of liquid formulations; 5) the firstpharmaceutical composition and/or second pharmaceutical compositionand/or liquid carrier further comprises an antioxidant, a bufferingagent, an acidifying agent, a solubilizing agent or solubility enhancingagent, a complexation enhancing agent, lyophilizing aids (for example,bulking agents or stabilizing agents), an electrolyte, anothertherapeutic agent, an alkalizing agent, an antimicrobial agent, anantifungal agent, a viscosity modifying agent, a flavoring agent, asweetening agent or a combination thereof; 6) the kit is providedchilled; 8) the liquid carrier and/or chamber has been purged with apharmaceutically acceptable inert gas to remove substantially all of theoxygen dissolved in the liquid carrier; 9) the chambers aresubstantially free from oxygen; 10) the liquid carrier further comprisesa buffering agent capable of maintaining a pH of about 2-7; 11) thechambers and solutions are sterile.

The term “unit dosage form” is used herein to mean a single dosage formcontaining a quantity of the active ingredient and the diluent orcarrier, said quantity being such that one or more predetermined unitsare normally required for a single therapeutic administration. In thecase of multi-dose forms, such as liquid-filled bottles, saidpredetermined unit will be one fraction such as a half or quarter of themultiple dose form. It will be understood that the specific dose levelfor any patient will depend upon a variety of factors including theindication being treated, therapeutic agent employed, the activity oftherapeutic agent, severity of the indication, patient health, age, sex,weight, diet, and pharmacological response, the specific dosage formemployed and other such factors.

The phrase “pharmaceutically acceptable” is employed herein to refer tothose compounds, materials, compositions, and/or dosage forms which are,within the scope of sound medical judgment, suitable for use in contactwith the tissues of human beings and animals without excessive toxicity,irritation, allergic response, or other problem or complication,commensurate with a reasonable benefit/risk ratio.

As used herein, the term “patient” or “subject” is taken to mean warmblooded animals such as mammals, for example, cats, dogs, mice, guineapigs, horses, bovine cows, sheep, non-humans, and humans.

The liquid formulation of the invention will comprise an effectiveamount of clopidogrel. By the term “effective amount”, it is understoodthat a therapeutically effective amount is contemplated. Atherapeutically effective amount is the amount or quantity ofclopidogrel that is sufficient to elicit the required or desiredtherapeutic response, or in other words, the amount that is sufficientto elicit an appreciable biological response when administered to asubject.

The invention also provides a pharmaceutical solid dosage formcomprising a therapeutically effective amount of clopidogrel, sulfoalkylether cyclodextrin (e.g. SBE7-β-CD), and a pharmaceutically acceptablesolid carrier, wherein the molar ratio of SAE-CD to clopidogrel is asdefined herein. The SAE-CD and clopidogrel can be included in the dosageform in admixture, as a preformed complex, or a combination thereof. Asolid dosage form can be prepared using conventional methods in thepharmaceutical sciences. In admixture, solid SAE-CD and solidclopidogrel are physically mixed using a low energy mixer to minimizecomplex formation. A solid preformed complex can be made by mixingSAE-CD and clopidogrel in a liquid carrier, such as an aqueous liquidcarrier, to form the complex, followed by removal of the liquid carrierthereby forming the solid preformed complex. Alternatively, the complexcan be prepared using a high energy milling process with the SAE-CD andclopidogrel, wherein either one or both of the two components isindependently provided in solid or liquid form.

The molar ratio of SAE-CD to clopidogrel in a composition or formulationof the invention will vary according to the pH of the formulation or thepH of an intended environment of use for a formulation. For example, themolar ratio can be less than about 6:1 when the pH of the formulation orof the intended environment of use is less than about 3.5.Alternatively, the molar ratio of SAE-CD to clopidogrel can be in therange of at least about 6:1, about 6:1 to 8:1, or at least about 8:1when the pH of the formulation or the intended environment of use isabout or greater than about 3.5. When the pH of the formulation orintended environment of use is about 5.5 or about 5.5 to 8, the molarratio can be at least about 6.5: 1, or at least 6.6:1. When the pH ofthe formulation or intended environment of use is about 8, the molarratio can be at least about 7.25:1, or at least 7.3:1.

The intended environment of use can be a pharmaceutical formulation orpart of a subject, such as part of a subject's circulatory system ordigestive tract, to which the formulation is administered. If theintended environment of use is the gastric region (meaning that theSAE-CD and clopidogrel is delivered to and released in the gastricregion such as the stomach or duodenum), then a molar ratio of less than6:1 or less than 3:1 can be used. If the intended environment of use isimmediately downstream of the gastric region (meaning that the SAE-CDand clopidogrel is delivered to and released in the jejunum or ileum),then a molar ratio of at least 6:1 or at least 8:1 can be used.

For oral administration and gastric delivery and release, the molarratio of SAE-CD, e.g. SBE7-β-CD, to clopidogrel can be reduced furtherto about 4:1 or less as is described in Examples 16 and 18 or to about0.5:1 or less as in Example 23 because either: 1) the pH of the GI tractaffects an increase in intrinsic solubility of the clopidogrel, therebyrequiring a lower molar ratio of SAE-CD to clopidogrel for dissolutionof the drug; or 2) the pH of the formulation can be at least mildlyacidic. The pH of the exemplary reconstitutable formulations in Examples16 and 18 ranges from pH 1-2.

In order for a liquid formulation of the invention to be clear, themolar ratio of SAE-CD to clopidogrel may vary according to the pH of theformulation. At a pH of about 8, the molar ratio should be at leastabout 7.25:1 or at least about 7.3:1, and at a pH of about 5.5, themolar ratio should be at least about 6.5:1 or at least about 6.6:1. Thismolar ratio is sufficient to provide a clear solution; however, highermolar ratios will result in improved stability against hydrolysis,photolysis, and chiral inversion by increasing the percentage ofclopidogrel bound by SAE-CD.

The SAE-CD to clopidogrel molar ratio formulations prepared according tothe examples below was determined. The pH of the liquid formulations wasalso determined. The data is summarized below.

Approx. Molar Example No. pH of solution Ratio 2 and 3 ~5.5 36:1 4 and 5~8.0 36.1 10, 11, 12 and 13 ~5.5  9:1 15 and 17 ~1.8 10:1 16 and 18 ~1.5and ~1.8  5:1 19 ~5.5 33:1 20 ~5.5 16:1 23 tablet  1:1 24 and 25 ~5.510:1

There is no need to provide an upper limit to the molar ratio of SAE-CDto clopidogrel, since excess SAE-CD present will only serve to stabilizethe clopidogrel further. However, the invention includes embodimentswherein the upper limit of the molar ratio of SAE-CD to clopidogrel isless than 1000:1, less than or about 500:1, less than or about 250:1,less than or about 100:1, less than or about 75:1, less than or about50:1, less than or about 40:1, less than or about 30:1, less than orabout 20:1, less than or about 17.5:1, less than or about 15:1, lessthan or about 12.5:1, less than or about 10:1. These upper limits can beused in combination with the other lower limits set forth herein.

The molar ratio of SAE-CD to clopidogrel for a composition or method ofthe invention can be within the range of 6:1 and 8:1, 6:1 to 10:1, 5:1to 12:1, 4:1 to 15:1, 5:1 to 14:1, 6:1 to 13:1, or 6:1 to 12.5:1; and/orat least 0.05:1, at least 0.25:1, at least 0.2:1, at least 0.5:1, atleast 1:1, at least 2:1, at least 3:1, at least 4:1, at least 5:1, atleast 6:1, at least 7:1, or at least 8:1; and/or at most 100:1, at most75:1, at most 50:1, at most 40:1, at most 35:1, at most 30:1, at most25:1, at most 20:1, at most 15:1, at most 14:1 at most 12.5:1, at most12:1, at most 10:1, at most 8:1, at most 6:1, at most 5:1, at most 4:1,at most 3:1, or at most 2:1.

Tablets prepared according to Example 23 were subject to dissolution inone liter of citric acid/phosphate buffer. PLAVIX® tablets were used ascontrol/reference samples. Each tablet contained 75mg free baseequivalents of clopidogrel. As can be seen in FIG. 9, both the preformedcomplex and physical mixture based tablets containing SAE-CD have afaster dissolution rate than the PLAVIX® tablets, and they also have ahigher final concentration. This faster dissolution time may lead to afaster absorption and hence faster/quicker therapeutic onset.

The typical oral daily dose for clopidogrel, expressed as the free base,is 75 mg. As a loading dose for certain minimally invasiveinterventional procedures, such as stent placement, angioplasty,intravascular ultrasound, atherectomy, carotid artery balloonangioplasty and stenting (CBAS), laser thrombolysis, brachytherapy,ultrasonography, use of intraarterial suction devices, use of snares, oruse of clot-retrieval devices, higher doses can be administered, e.g.900 mg, 750 mg, 675 mg, 600 mg, 450 mg, 375 mg, 300 mg, 225 mg, 200 mg,150 mg, 100 mg, 75 mg, 50 mg, 40 mg, 30 mg, 25 mg, 20 mg, 15 mg, 12.5mg, 10 mg, 7.5 mg, 5 mg, 2 mg, 1 mg, 0.75 mg, or 0.1 mg of clopidogrel,or in the range of 0.1 to 900 mg, 0.1 to 100 mg, 100 to 300 mg, about300 mg, 300 to 600 mg, 300 to 900 mg, 600 to 900 mg, 50 to 600 mg, 75 to600 mg, 150 to 600 mg, or 200 to 450 mg of clopidogrel. Althoughtitration of the dose is not indicated for PLAVIX®, the presentformulation is easily divisible to facilitate careful dose titration ifneeded. The present formulations can be administered as directed in thepackage insert for the PLAVIX® tablet formulation, meaning a subject canbe administered a dose of the present formulation containing anequivalent amount of clopidogrel, which would be about 75, about 300 orabout 75 to about 300 mg. The Physician's Desk Reference 56^(th) ed.(pp. 3084-3086; Eds. Lori Murray, Gwynned L. Kelly; Medical EconomicsCompany, Inc., Montvale, N.J. 07645-1742, 2002), the relevant text ofwhich is hereby incorporated by reference, discloses the package insertfor PLAVIX®, and particularly the dosage and administration for theformulation.

As used herein, the term “bleeding time” refers to the amount of time ittakes for bleeding to stop in a mammal after controlled, standardizedpuncture of the skin of the mammal, for example, after controlled,standardized puncture of the earlobe or forearm of the mammal (orsubject). Bleeding is stopped in part by platelet aggregation in vivo;therefore, bleeding time increases as the rate or overall amount ofplatelet aggregation decreases, and bleeding time decreases as the rateor overall amount of platelet aggregation increases. The relationshipbetween bleeding time and platelet aggregation is not necessarilylinear. Bleeding time is an indirect measure of the extent of orpotential for platelet aggregation.

A bleeding time assay can be conducted according to standardized assaysdescribed in the literature (Sramek, et al, Thrombosis and Haemostasis67 (5) 514-518 (1992)) or according to Example 21, wherein the bleedingtime is measured after controlled, standardized puncture of the skin ofa subject. A bleeding time assay measures the time it takes for a smallblood vessel to close off and stop bleeding after it has been punctured.

The percentage increase in a subject's bleeding time is determined byfirst measuring the subject's “baseline bleeding time” (BBT, sec) priorto administration of a formulation or composition of the invention. Thesubject is then administered a formulation or composition of theinvention, and the subject's “treatment bleeding time” (TBT, sec) ismeasured. The percentage increase (δBT, %) is then calculated asfollows:

δBT=((TBT−BBT)/BBT)*100

Known platelet aggregation assays and standard point-of-care equipmentcan be employed to determine the extent of or rate of (or potential for)platelet aggregation ex vivo. Most determinations are performed usingeither an optical-based method or an impedance-based method(Dyszkiewicz-Korpanty, et al., Clinical and AppliedThrombosis/Hemostasis, Vol. 11, No. 1, 25-35 (2005), the entiredisclosure of which is hereby incorporated by reference). Exemplaryequipment, systems and assays include a platelet aggregometer(Chrono-log Corporation, Havertown, Pa.), VerifyNow PlateletAggregometry (Accumetrics, San Diego, Calif.), Platelet AggregationProfiler™ (Bio/Data Corporation, Horsham, Pa.) Slide PlateletAggregation Test™ (SPAT™, Analytical Control Systems, Inc., Fishers,Ind.), Heparin Induced Platelet Aggregation test, Ristocetin TitrationPlatelet Aggregation assay, and others. The assay is generally basedupon impedance or optical aggregometry. In general, a sample of asubject's blood is obtained and the plasma is separated from the sample.The plasma is treated with ADP, as described herein, and the extent ofplatelet aggregation in the plasma is then determined. The assay can beconducted before or after clopidogrel is administered to a subject.

The percentage decrease in a subject's extent of (or potential for)platelet aggregation is determined by first measuring the subject's“baseline platelet aggregation” (BPA) prior to administration of aformulation or composition of the invention. The subject is thenadministered a formulation or composition of the invention, and thesubject's “treatment platelet aggregation” (TPA) is measured. Thepercentage decrease (δPA, %) is then calculated as follows:

δPA=((BPA−TPA)/BPA)*100

An in vivo evaluation in mice of a clear liquid formulation, preparedaccording to Example 20, was conducted according to Example 21. Thebleeding time of the individual mice was measured periodically todetermine the difference in performance between a PLAVIX® formulation(oral, ground tablet), clopidogrel bisulfate-SAE-CD (i.v.), clopidogrelbisulfate-SAE-CD (oral, solution). A reduction in the time to achieve anincreased bleeding time is considered a reduction in the time totherapeutic onset, since an increase in bleeding time marks thebeginning of therapeutic onset. The results (FIG. 8) indicate that useof a formulation according to the invention results in a more rapidtherapeutic onset, whereby the amount of time after administrationrequired to achieve a desired increase in bleeding time is substantiallyreduced as compared to administration of the PLA VIX® solid(administered as a powder prepared by crushing or grinding a tablet.)The formulation of the invention provides a more rapid therapeutic onsetwithout requiring such excessive doses as are currently administered forinterventional cardiology procedures.

Surprisingly, the oral dose of the invention can provide almost the samerate of therapeutic onset and level of therapeutic effect as does theparenteral i.v. dose of the invention, and both provide a substantiallymore rapid onset of therapeutic benefit and a higher level oftherapeutic effect than does a reference formulation, for example anoral tablet, containing an equivalent dose of clopidogrel.

Related to the reduced time to therapeutic onset, the invention providesa reduced time to peak or target therapeutic effect. In someembodiments, the invention provides a method of decreasing the time topeak or target therapeutic effect in a responder subject administeredclopidogrel, the method comprising: administering to a subject in needthereof a first composition comprising SAE-CD and a therapeuticallyeffective amount of clopidogrel sufficient to achieve a targettherapeutic effect, whereby the time to peak or target therapeuticeffect achieved by administration of the first composition is less thanthe time to peak therapeutic effect achieved by similar administrationof an otherwise similar reference composition, excluding SAE-CD, andcomprising substantially the same therapeutically effective amount ofclopidogrel. The reference composition can be a tablet. The firstcomposition can be a solid, suspension or liquid.

FIGS. 10 to 12 are log-linear plots of mean plasma concentration ofcompound versus time after administration for the compounds clopidogrel(FIG. 10), the thiol metabolite of clopidogrel (FIG. 11) and thecarboxylic acid metabolite of clopidogrel (FIG. 12). The data wereobtained as part of the clinical study detailed in Example 27. FIG. 10depicts a clear dose to plasma concentration relationship forclopidogrel administered parenterally, i.e. intravenously, for 0.1, 1.0,10, 30, 100 and 300 mg doses. The higher the dose administered, thehigher the plasma concentration of clopidogrel. The time to peak plasmaconcentration was about 30 sec or about 1-5 minutes, or less than orabout 10-15 minutes, and greater than or about the end of theadministration period or by termination of administration. The time topeak plasma concentration can be affected by the rate of parenteraladministration: the longer the period of administration, the longer thetime to peak plasma concentration. As noted below, the clopidogrel wasadministered i.v. over as a bolus over a period of less than or about 1min (for samples comprising 0.1 mg, 1 mg, 10 mg, or 30 mg ofclopidogrel), about 3 to 5 minutes or about 4 minutes (for samplescomprising about 100 mg of clopidogrel), and about 6 to 10 min or about8 minutes (for samples comprising about 300 mg of clopidogrel).

FIG. 11 depicts a dose to plasma concentration relationship for thethiol metabolite of clopidogrel. The data were obtained as part of theclinical study detailed in Example 27. In general, the higher the doseof clopidogrel, the higher the corresponding plasma concentration of themetabolite, which is thought to be the pharmacologically activemetabolite. It is important to note that low doses of clopidogrel canprovide a therapeutic response even though the corresponding plasmaconcentration of the thiol metabolite may be below its detection limit,at least according to the assay used herein. Accordingly, it is notnecessary, for the methods of the invention, that the thiol metabolitebe quantifiable or even detectable. It is only necessary that the amountof clopidogrel converted in vivo to the thiol metabolite is sufficientto provide a corresponding therapeutic effect in a subject within aspecified period of time.

FIG. 12 depicts a dose to plasma concentration relationship for thecarboxylic acid metabolite of clopidogrel. The data were obtained aspart of the clinical study detailed in Example 27. In general, thehigher the dose of clopidogrel, the higher the corresponding plasmaconcentration of the metabolite, which is thought to be apharmacologically inactive metabolite.

FIG. 13 depicts mean dose-response curves for the subjects administeredclopidogrel according to Example 27. The response was determined aspercent of platelet aggregation inhibition following exposure of thesubjects' plasma ex-vivo to ADP (5 βM). In general, the time to onset ofthe therapeutic effect was less than or about 15 minutes. As describedby Weerakody et al. 2007 (see previous citation), maximal plateletaggregation inhibition of >=15% is a recognized therapeutic effect todifferentiate various anti-platelet drugs known to be useful in thetreatment of ACS.

As a result of the more rapid therapeutic onset, more rapid time to peakplasma concentration, more rapid time to peak therapeutic effect, morerapid time to target therapeutic effect provided by the methods andcompositions of the invention, several clinical advantages are providedby the same as compared to pharmaceutical oral tablet compositions ofclopidogrel and administration and uses thereof. For instance, thecompositions and methods of the invention provide reduced treatmenttime, reduced immediate risk of occurrence of a second cardiovascularevent in a subject by reducing treatment/response time, such as byreducing the time to peak therapeutic, time to peak plasmaconcentration, time to target therapeutic effect, and time totherapeutic onset.

FIG. 14 depicts a plot of the percentage of responder subjects versusthe time period after administration of clopidogrel according to Example27. As used herein a “responder” subject is a subject that has beentreated with clopidogrel and whose plasma exhibits at least a 15%platelet aggregation inhibition when the subjects' plasma is exposed exvivo to 5 βM ADP and the extent of platelet aggregation is determined byplatelet aggregometry. Any known method of platelet aggregometry can beused according to the invention; however, suitable methods are describedherein. In general, the higher the dose administered to a group ofsubjects, the higher the percentage of responder subjects identified inthat group. As well, the higher the dose administered to a group ofsubjects, the more rapid the group reached its maximum number ofresponder subjects. A composition or formulation of the invention can beused to treat, prevent, ameliorate, reduce the occurrence of, or reducethe risk of occurrence of a disease, disorder or condition that istherapeutically responsive to clopidogrel therapy. As used herein asregards a method of treatment of a subject, the term “treat” or“treating” means to alleviate, ameliorate, eliminate, reduce theseverity of, reduce the frequency of, occurrence of, or prevent symptomsassociated with a disease, disorder or condition having excessive orundesired platelet aggregation as an etiological component. As usedherein, the term “therapeutically responsive to clopidogrel” means thattreatment of a subject with such a disease, disorder or condition with atherapeutically effective amount of clopidogrel will result in aclinical benefit or therapeutic benefit in the subject. The method oftreating, preventing, ameliorating, reducing the occurrence of, orreducing the risk of occurrence of a disease, disorder or condition thatis therapeutically responsive to clopidogrel therapy in a subjectcomprises administering to the subject in need thereof a formulation orcomposition of the invention, wherein the formulation or compositioncomprises SAE-CD and a dose of clopidogrel. A therapeutically effectiveamount of clopidogrel can include one, two, or more doses ofclopidogrel.

In some embodiments, the disease, disorder or condition includes athrombotic disease, disorder, or condition and can be selected from thegroup consisting of myocardial infarction, stroke, establishedperipheral arterial disease (PAD), secondary ischemic events, acutecoronary syndrome (ACS, e.g. unstable angina/non-Q-wave MI), transientischemic attack, cerebral arteriosclerosis, cerebrovascular disease,cardiovascular disease, angina pectoris, deep vein thrombosis, pulmonaryemboli (PE), sickle cell crisis, and cardiac arrhythmia.

The compositions, methods, and formulations of the invention can beemployed in both interventional and non-interventional treatmentprotocols for subjects presenting with a cardiovascular condition suchas, ACS (Acute Coronary Syndrome), which refers to any group of clinicalsymptoms compatible with acute myocardial ischemia. Acute myocardialischemia is chest pain due to insufficient blood supply to the heartmuscle that results from coronary artery disease. They can be employedin medical treatment protocols requiring a minimally invasiveinterventional procedure such as PCI (stent replacement or balloonangioplasty) or in medical treatment protocols not requiring invasiveprocedures. In order to benefit from clopidogrel therapy, the subjectwould necessarily be a responder, as defined herein. The subject may ormay not be undergoing chronic clopidogrel therapy prior to presentingwith the ACS. In its broadest sense, the treatment protocol comprisesadministering to the subject in need thereof a pharmaceuticalcomposition comprising SAE-CD and clopidogrel in an amount sufficient toprovide a target therapeutic effect in the subject within a period of 10sec to 120 min, 30 sec to 120 min, 30 sec to 100 min, 30 sec to 90 min,30 sec to 60 min, 1 min to 60 min, 1 min to 45 min, 1 min to 30 min, 1min to 20 min, or 1 min to 15 min, 1 min to 90 min, 5 min to 60 min, 10min to 60 min, 5 min to 45 min, 10 min to 45 min, 15 min to 30 min, 5min to 30 min, 5 min to 15 min, or 10 min to 20 min, or within less thanabout 10 seconds, less than about 1 min, less than about 2.5 min, lessthan about 5 min, less than about 7.5 min, less than about 10 min, lessthan about 15 min, less than about 20 min, less than about 30 min, lessthan about 120 min, less than about 100 min, less than about 90 min,less than about 75 min, less than about 60 min, less than about 50 min,less than about 45 min, or less than about 40 min after administrationof the composition.

In this regard, the target therapeutic effect is that level of plateletaggregation inhibition or bleed time as deemed sufficient by a clinicianto permit performance of the minimally invasive interventionalprocedure, such as PCI. The target therapeutic effect may vary fromsubject to subject according to their ability to absorb and/ormetabolize clopidogrel to its active metabolite. The therapeutic effectmay vary according to the disease, disease, disorder or condition beingtreated; however, any and all clinically beneficial therapeutic effectsachieved in a subject as a result of the in vivo inhibition of plateletaggregation or increased bleeding time caused by clopidogrel, whenadministered to the subject in the compositions and/or according to themethods of the invention, are contemplated.

The treatment protocol can further comprise the earlier step ofdetermining whether or not the subject requires interventional ornon-interventional medical treatment. Following administration of theclopidogrel, the subject may undergo the minimally invasive procedure inas little as 10 sec to 120 min, 30 sec to 120 min, 30 sec to 100 min, 30sec to 90 min, 30 sec to 60 min, 1 min to 60 min, 1 min to 45 min, 1 minto 30 min, 1 min to 20 min, or 1 min to 15 min, 1 min to 90 min, 5 minto 60 min, 10 min to 60 min, 5 min to 45 min, 10 min to 45 min, 15 minto 30 min, 5 min to 30 min, 5 min to 15 min, or 10 min to 20 min, orwithin less than about 10 seconds, less than about 1 min, less thanabout 2.5 min, less than about 5 min, less than about 7.5 min, less thanabout 10 min, less than about 15 min, less than about 20 min, less thanabout 30 min, less than about 120 min, less than about 100 min, lessthan about 90 min, less than about 75 min, less than about 60 min, lessthan about 50 min, less than about 45 min, or less than about 40 minafter parenteral or peroral administration of the clopidogrel-containingcomposition of the invention.

For a treatment protocol wherein a non-interventional procedure (medicaltreatment) and clopidogrel administration are indicated for the subject,the subject will be administered clopidogrel in a composition of theinvention and a second therapeutic agent, such as described herein.

In some embodiments, the invention provides a treatment protocol for asubject presenting with a cardiovascular condition, disease or disorder,the method comprising: a) determining whether or not the subjectrequires interventional or non-interventional medical treatment; and b)if the subject requires minimally invasive interventional medicaltreatment, then administering to the subject a pharmaceuticalcomposition comprising SAE-CD and clopidogrel in an amount sufficient toprovide a target therapeutic effect in the patient within a period of 10sec to 120 min, 30 sec to 120 min, 30 sec to 100 min, 30 sec to 90 min,30 sec to 60 min, 1 min to 60 min, 1 min to 45 min, 1 min to 30 min, 1min to 20 min, or 1 min to 15 min, 1 min to 90 min, 5 min to 60 min, 10min to 60 min, 5 min to 45 min, 10 min to 45 min, 15 min to 30 min, 5min to 30 min, 5 min to 15 min, or 10 min to 20 min, or within less thanabout 10 seconds, less than about 1 min, less than about 2.5 min, lessthan about 5 min, less than about 7.5 min, less than about 10 min, lessthan about 15 min, less than about 20 min, less than about 30 min, lessthan about 120 min, less than about 100 min, less than about 90 min,less than about 75 min, less than about 60 min, less than about 50 min,less than about 45 min, or less than about 40 min after saidadministration, and conducting the minimally invasive interventionalprocedure; or c) if the subject requires non-interventional medicaltreatment, then administering to the subject a pharmaceuticalcomposition comprising SAE-CD and clopidogrel in an amount sufficient toprovide a target therapeutic effect in the subject within a period of 10sec to 120 min, 30 sec to 120 min, 30 sec to 100 min, 30 sec to 90 min,30 sec to 60 min, 1 min to 60 min, 1 min to 45 min, 1 min to 30 min, 1min to 20 min, or 1 min to 15 min, 1 min to 90 min, 5 min to 60 min, 10min to 60 min, 5 min to 45 min, 10 min to 45 min, 15 min to 30 min, 5min to 30 min, 5 min to 15 min, or 10 min to 20 min, or within less thanabout 10 seconds, less than about 1 min, less than about 2.5 min, lessthan about 5 min, less than about 7.5 min, less than about 10 min, lessthan about 15 min, less than about 20 min, less than about 30 min, lessthan about 120 min, less than about 100 min, less than about 90 min,less than about 75 min, less than about 60 min, less than about 50 min,less than about 45 min, or less than about 40 min and providing to thesubject said non-interventional medical treatment; or d) if the subjectrequires invasive interventional medical treatment (e.g. CABG), then notadministering to the subject clopidogrel. In some embodiments, theprotocol comprises: 1) optionally sedating patient presenting; 2)determining for high risk of ACS; 3) optionally, alerting the catheterlab of incoming patient; 4) optionally, transporting patient to thecatheter lab; 5) performing one or more diagnostic coronary tests on thepatient; 6) determining if PCI or CABG is indicated; and 7) if PCI isindicated, parenterally administering a liquid composition orformulation comprising clopidogrel, and optionally SAE-CD, or perorallyadministering a composition or formulation comprising clopidogrel andSAE-CD, performing PCI, and optionally maintaining the patient on longterm (chronic) clopidogrel therapy; or 8) if CABG is indicated,performing the CABG, without administration of clopidogrel. The methodof the invention can include additional steps as needed, such as anystep(s) indicated by an attending clinician. The method of sedating caninclude administration of an anxiolytic agent, NSAID, non-narcotic painrelieving agent, narcotic pain relieving agent, anesthetic, sedative,and/or anti-inflammatory agent.

Suitable diagnostic coronary tests include coronary angiography,electrocardiography, EKG, echocardiography, CT scanning angiography andany other test knows to those of skill in the art for determiningcoronary dysfunction the result of which might require an interventionalor non-interventional treatment.

A non-interventional procedure is defined as a non-surgical procedure. Anon-interventional procedure can be administration of one or more othertherapeutic agents to the subject or another non-surgical procedure asdescribed herein. An interventional procedure is defined as an invasiveor minimally invasive surgical procedure. An invasive procedure can beCABG. A minimally invasive procedure can be PCI.

Given that clopidogrel therapy will only be useful to a respondersubject, the invention provides a method of rapidly determining whethera subject is a responder or a non-responder. The method comprises:administering to the subject a composition comprising an “expectedtherapeutically effective amount” of clopidogrel, and determining thesubject's responsiveness to the clopidogrel within a period of 10 sec to120 min, 30 sec to 120 min, 30 sec to 100 min, 30 sec to 90 min, 30 secto 60 min, 1 min to 60 min, 1 min to 45 min, 1 min to 30 min, 1 min to20 min, or 1 min to 15 min, 1 min to 90 min, 5 min to 60 min, 10 min to60 min, 5 min to 45 min, 10 min to 45 min, 15 min to 30 min, 5 min to 30min, 5 min to 15 min, or 10 min to 20 min, or within less than about 10seconds, less than about 1 min, less than about 2.5 min, less than about5 min, less than about 7.5 min, less than about 10 min, less than about15 min, less than about 20 min, less than about 30 min, less than about120 min, less than about 100 min, less than about 90 min, less thanabout 75 min, less than about 60 min, less than about 50 min, less thanabout 45 min, or less than about 40 min after administration of thecomposition to the subject. By “expected therapeutically effectiveamount” is meant that amount of clopidogrel that would be expected toprovide a therapeutic effect in the subject at the time the subject isadministered the composition. An expected therapeutically effectiveamount is generally 900 mg, 750 mg, 675 mg, 600 mg, 450 mg, 375 mg, 300mg, 225 mg, 200 mg, 150 mg, 100 mg, 75 mg, 50 mg, 40 mg, 30 mg, 25 mg,20 mg, 15 mg, 12.5 mg, 10 mg, 7.5 mg, 5 mg, 2 mg, 1 mg, 0.75 mg, or 0.1mg of clopidogrel, or in the range of 0.1 to 900 mg, 0.1 to 100 mg, 100to 300 mg, about 300 mg, 300 to 600 mg, 300 to 900 mg, 600 to 900 mg, 50to 600 mg, 75 to 600 mg, 150 to 600 mg, or 200 to 450 mg of clopidogrel.

According to the method, a responder subject is one that exhibits >/=15%inhibition of platelet aggregation, as determined with an ex vivo assay,following administration of an expected therapeutically effective amountof a composition or formulation of the invention. A non-responder(otherwise known as a poor responder, hypo-responder, intermediateresponder, low responder or resistant) subject is one in which <15%inhibition of platelet aggregation, as determined with an ex vivo assay,following administration of an expected therapeutically effective amountof a composition or formulation of the invention. Platelet aggregationinhibition is defined as the % of platelet aggregation induced by 5uMadenosine diphosphate (ADP) in an ex vivo assay. Suitable methods foruse in defining a subject as a responder versus as non-responder aredisclosed by Weerakkody et al. (J. Cardiovascular Pharmacol. Therap.(2007), 12(3), 205-212), the entire disclosure of which is herebyincorporated by reference.

The method of the invention is advantageously conducted in a shortertime period than can be achieved by an otherwise similar method wherethe clopidogrel is administered as a solid oral tablet. In other words,parenteral or peroral administration to a subject of clopidogrel in acomposition or formulation of the invention will provide a faster onsetof a therapeutic response in a responder subject than does peroraladministration of a solid oral dosage form, e.g. PLAVIX®, tablet(s), notof the invention but comprising substantially the same amount ofclopidogrel. This means the time period, within which a clinician isable to determine whether or not a subject is a responder, will beshorter when the subject is administered a composition of the inventionthan when the subject is orally administered a solid dosage form not ofthe invention, e.g. PLAVIX®. The time period for such determinationaccording to the invention is generally in the range of 10 sec to 120min, 30 sec to 120 min, 30 sec to 100 min, 30 sec to 90 min, 30 sec to60 min, 1 min to 60 min, 1 min to 45 min, 1 min to 30 min, 1 min to 20min, or 1 min to 15 min, 1 min to 90 min, 5 min to 60 min, 10 min to 60min, 5 min to 45 min, 10 min to 45 min, 15 min to 30 min, 5 min to 30min, 5 min to 15 min, or 10 min to 20 min, or within less than about 10seconds, less than about 1 min, less than about 2.5 min, less than about5 min, less than about 7.5 min, less than about 10 min, less than about15 min, less than about 20 min, less than about 30 min, less than about120 min, less than about 100 min, less than about 90 min, less thanabout 75 min, less than about 60 min, less than about 50 min, less thanabout 45 min, or less than about 40 min after administration of thecomposition.

The step of determining the subject's responsiveness to clopidogrel cancomprise: obtaining a sample of blood of the subject; and determiningthe extent of platelet aggregation in the subject's plasma. Suitablemethods for determining the extent of platelet aggregation in thesubject's plasma include, by way of example and without limitation,aggregometry, such as light transmittance or impedance aggregometry.

The composition and formulation of the invention provides one or moreadvantages over peroral administration of clopidogrel-containingtablets, e.g. PLAVIX®. One such advantage is dose reduction. A subjectbeing perorally or parenterally administered a dose of clopidogrel in acomposition or formulation of the invention will require a firsttherapeutically effective amount in order to receive a therapeuticbenefit from the clopidogrel. The same subject being perorallyadministered a dose of clopidogrel in a tablet dosage form, notaccording to the invention, will require a second therapeuticallyeffective amount in order to receive substantially the same therapeuticbenefit from the clopidogrel. For that subject the first therapeuticallyeffective amount can be and is generally less than the secondtherapeutically effective amount. Accordingly, a method of reducing therequired therapeutic dose in a responder subject in need of clopidogreltherapy, the method comprising: parenterally or perorally administeringto the subject a first therapeutically effective amount of clopidogrelin a pharmaceutical composition comprising SAE-CD, wherein the firsttherapeutically effective amount is at least 1.1-fold, at least1.2-fold, at least 1.25-fold, at least 1.5-fold, at least 2-fold, atleast 3-fold, at least 4-fold, at least 5-fold, at least 7-fold, atleast 8-fold, at least 10-fold smaller, at least 15-fold, at least20-fold, about 1.1 to about 20-fold, about 1.2-fold to about 15-fold,about 1.25-fold to about 10-fold, about 2-fold to about 10-fold, orabout 3 fold to about 8-fold smaller than a second therapeuticallyeffective amount, which is the amount of clopidogrel required to providesubstantially the same therapeutic effect when clopidogrel isadministered to the subject perorally in a reference solidpharmaceutical composition excluding SAE-CD.

The parenteral administration of clopidogrel to a subject in needthereof according to the invention may provide an alternative means oftherapy for a subject that is a non-responder in terms of peroraladministration of clopidogrel. For example, it may occur that a subjectbeing perorally administered a tablet comprising clopidogrel (such as aPLAVIX tablet) will not exhibit a sufficiently high therapeutic responseto be deemed a “responder”, as defined herein. However, that samesubject may become a responder by being administered the clopidogrelparenterally. In one clinical study, a group of subjects wereadministered clopidogrel (10-300 mg) perorally, and those subjects didnot exhibit a sufficiently high therapeutic response to be deemed“responders”. Clopidogrel was then administered parenterally to thosesame subjects, after which 9-25% of the subjects exhibited asufficiently high therapeutic response to be deemed “responders”.Accordingly, the invention provides a method of converting anon-responder subject, in terms of peroral administration ofclopidogrel, to a responder subject, the method comprising parenterallyadministering clopidogrel to the subject in need thereof, therebyproviding a therapeutic response to clopidogrel in the subject, whereinthe therapeutic response is sufficient to deem the subject a responder.

Another such advantage is the ability of a clinician to titrate the doseadministered to a subject within a shorter period of time than iscurrently possible in clopidogrel therapy in order to achieve a targettherapeutic effect in the subject. A composition or formulationcomprising an amount of clopidogrel is administered to a subject, andthe corresponding therapeutic effect in the subject is determined withina specified period of time. That period of time will be shorter when theclopidogrel is administered parenterally as a liquid dosage form thanwhen it is administered perorally as a tablet dosage form, e.g. PLAVIX®,tablet. The importance of this advantage relies upon the acceptedclinical target to get a patient presenting with ACS from entry into anemergency room or operating room to initiation of a PCI procedure within90 minutes, specifically doing so while not precluding the utility ofCABG by not treating with PLAVIX prior to coronary angiography.

Accordingly, the invention provides a method of escalating dose in asubject to achieve a target therapeutic effect in the subject, themethod comprising: parenterally administering to the subject an amountof clopidogrel; within a delay period of less than 10 sec to 120 min, 30sec to 120 min, 30 sec to 100 min, 30 sec to 90 min, 30 sec to 60 min, 1min to 60 min, 1 min to 45 min, 1 min to 30 min, 1 min to 20 min, or 1min to 15 min, 1 min to 90 min, 5 min to 60 min, 10 min to 60 min, 5 minto 45 min, 10 min to 45 min, 15 min to 30 min, 5 min to 30 min, 5 min to15 min, or 10 min to 20 min, or within less than about 10 seconds, lessthan about 1 min, less than about 2.5 min, less than about 5 min, lessthan about 7.5 min, less than about 10 min, less than about 15 min, lessthan about 20 min, less than about 30 min, less than about 120 min, lessthan about 100 min, less than about 90 min, less than about 75 min, lessthan about 60 min, less than about 50 min, less than about 45 min, orless than about 40 min after the parenteral administration, determiningthe corresponding therapeutic effect achieved in the subject; and if theextent of therapeutic effect achieved is less than the targettherapeutic effect, repeating the steps of parenterally administeringand determining until the target therapeutic effect is achieved. Thesteps can be repeated as many times as needed to achieve the targettherapeutic effect. The amount of clopidogrel administered willgenerally be in the range of about 50 to 600 mg, 0.1 to 900 mg, 1 to 900mg, 10 to 900 mg, 0.1 to 100 mg, 25 to 750 mg, 50 to 600 mg, 75 to 600mg, 75 to 500 mg, 100 to 300 mg, 100 to 400 mg, 150 to 600 mg, or 200 to450 mg, 200 to 400 mg, 300 to 600 mg, 300 to 900 mg, 600 to 900 mg,about 0.1 mg, about 0.75 mg, about 1 mg, about 2 mg, about 5 mg, about7.5 mg, about 10 mg, about 12.5 mg, about 15 mg, about 20 mg, about 25mg, about 30 mg, about 40 mg, about 50 mg, about 75 mg, about 100 mg,about 150 mg, about 200 mg about 225 mg, about 300 mg, about 375 mg,about 450 mg, about 525 mg, about 600 mg, about 675 mg, about 750 mg,about 900 mg. The delay period is generally 10 sec to 120 min, 30 sec to120 min, 30 sec to 100 min, 30 sec to 90 min, 30 sec to 60 min, 1 min to60 min, 1 min to 45 min, 1 min to 30 min, 1 min to 20 min, or 1 min to15 min, 1 min to 90 min, 5 min to 60 min, 10 min to 60 min, 5 min to 45min, 10 min to 45 min, 15 min to 30 min, 5 min to 30 min, 5 min to 15min, or 10 min to 20 min, or within less than about 10 seconds, lessthan about 1 min, less than about 2.5 min, less than about 5 min, lessthan about 7.5 min, less than about 10 min, less than about 15 min, lessthan about 20 min, less than about 30 min, less than about 120 min, lessthan about 100 min, less than about 90 min, less than about 75 min, lessthan about 60 min, less than about 50 min, less than about 45 min, orless than about 40 min. The clopidogrel can be present in an aqueousliquid composition or formulation comprising clopidogrel SAE-CD.

The step of determining can comprise: obtaining a sample of plasma ofthe subject; and determining the extent of platelet aggregation in thesubject's plasma by aggregometry. The step of obtaining can comprise:obtaining a sample of blood of the patient; and separating the plasmafrom the blood to form a plasma sample, then conducting plateletaggregometry.

The invention also provides a method of escalating dose in a subject toachieve a target therapeutic effect in the subject, the methodcomprising: perorally administering to the subject an amount ofclopidogrel in a composition or formulation comprising clopidogrel andSAE-CD; within a delay period of 10 sec to 120 min, 30 sec to 120 min,30 sec to 100 min, 30 sec to 90 min, 30 sec to 60 min, 1 min to 60 min,1 min to 45 min, 1 min to 30 min, 1 min to 20 min, or 1 min to 15 min, 1min to 90 min, 5 min to 60 min, 10 min to 60 min, 5 min to 45 min, 10min to 45 min, 15 min to 30 min, 5 min to 30 min, 5 min to 15 min, or 10min to 20 min, or within less than about 10 seconds, less than about 1min, less than about 2.5 min, less than about 5 min, less than about 7.5min, less than about 10 min, less than about 15 min, less than about 20min, less than about 30 min, less than about 120 min, less than about100 min, less than about 90 min, less than about 75 min, less than about60 min, less than about 50 min, less than about 45 min, or less thanabout 40 min after the peroral administration, determining thecorresponding therapeutic effect achieved in the subject; and if theextent of therapeutic effect achieved is less the target therapeuticeffect, repeating the steps of perorally administering and determininguntil the target therapeutic effect is achieved. The steps can berepeated as many times as needed to achieve the target therapeuticeffect. The amount of clopidogrel administered will generally be in therange of 50 to 600 mg, 0.1 to 900 mg, 1 to 900 mg, 10 to 900 mg, 0.1 to100 mg, 25 to 750 mg, 50 to 600 mg, 75 to 600 mg, 75 to 500 mg, 100 to300 mg, 100 to 400 mg, 150 to 600 mg, or 200 to 450 mg, 200 to 400 mg,300 to 600 mg, 300 to 900 mg, 600 to 900 mg, about 0.1 mg, about 0.75mg, about 1 mg, about 2 mg, about 5 mg, about 7.5 mg, about 10 mg, about12.5 mg, about 15 mg, about 20 mg, about 25 mg, about 30 mg, about 40mg, about 50 mg, about 75 mg, about 100 mg, about 150 mg, about 200 mgabout 225 mg, about 300 mg, about 375 mg, about 450 mg, about 525 mg,about 600 mg, about 675 mg, about 750 mg, about 900 mg. The delay periodis generally 10 sec to 120 min, 30 sec to 120 min, 30 sec to 100 min, 30sec to 90 min, 30 sec to 60 min, 1 min to 60 min, 1 min to 45 min, 1 minto 30 min, 1 min to 20 min, or 1 min to 15 min, 1 min to 90 min, 5 minto 60 min, 10 min to 60 min, 5 min to 45 min, 10 min to 45 min, 15 minto 30 min, 5 min to 30 min, 5 min to 15 min, or 10 min to 20 min, orwithin less than about 10 seconds, less than about 1 min, less thanabout 2.5 min, less than about 5 min, less than about 7.5 min, less thanabout 10 min, less than about 15 min, less than about 20 min, less thanabout 30 min, less than about 120 min, less than about 100 min, lessthan about 90 min, less than about 75 min, less than about 60 min, lessthan about 50 min, less than about 45 min, or less than about 40 min.

As used herein, the term “about” is taken to mean +/−10% of theindicated value.

In view of the above description and the examples below, one of ordinaryskill in the art will be able to practice the invention as claimedwithout undue experimentation. The foregoing will be better understoodwith reference to the following examples that detail certain proceduresfor the preparation of formulations according to the present invention.All references made to these examples are for the purposes ofillustration. The following examples should not be consideredexhaustive, but merely illustrative of only a few of the manyembodiments contemplated by the present invention.

EXAMPLE 1

The solubility of clopidogrel bisulphate was determined in −20% w/v ofHP-β-CD, SBE-γ-CD, Captisol® (SBE-β-CD), SBE-α-CD, SBE-α-CD, SBE-α-CD,SPE-α-CD, SPE-β-CD SPE-γ-CD, ADVASEP (SBE-β-CD) and α-CD with DS numbersof 7.6, 6.1, 6.6, 2.4, 3.9, 5.4, 5, 5.6, 5.4, 5.5 and 0 (unsubstituted),respectively. This was done according to procedures well known in theart (Higuchi et al. in Phase Solubility Techniques, in Advances inAnalytical Chemistry and Instrumentation (Ed. CN. Reilly, John Wiley &Sons Inc., Vol. 4 (1965), pg. 117-212) the relevant disclosure of whichis hereby incorporated by reference). The results are detailed in thetable below.

Cyclo- Molar dextrin Ratio of Molecular Cyclodextrin ClopidogrelClopidogrel Weight Concentration Solubilized to Cyclodextrin (g/mole)(M) (M) Cyclodextrin HP7.6-β-CD 1575.8 0.117 0.005 1:23 SBE6.1-γ-CD2260.8 0.074 0.006 1:12 SBE6.6-β-CD 2168 0.092 0.012 1:8 SBE2.4-α-CD1358 0.141 0.0004 1:353 SBE3.9-α-CD 1594 0.121 0.0008 1:151 SBE5.8-α-CD1896 0.100 0.0017 1:59 SPE5-α-CD 1699.2 0.100 0.001 1:100 SPE5.6-β-CD1945.72 0.076 0.005 1:15 SPE5.4-γ-CD 2074.6 0.090 0.001 1:90 SBE5.5-β-CD2004 0.082 0.011 1:8 α-CD 972 0.103 0.0004 1:258

EXAMPLE 2

An aqueous solution of clopidogrel bisulfate was prepared at pH 5.5. Theformulation comprised Captisol® (SBE-β-CD, DS=6.6) (39.0% wt./vol.) andclopidogrel bisulfate. The amounts used are specified in the tablebelow.

Ingredients Amount Clopidogrel Bisulfate 300.0 mg (equivalent to 229.9mg clopidogrel) SBE-β-CD 39.0 g (anhydrous basis) Sodium Phosphate,monobasic buffer 0.2M

The following procedure was used to prepare the formulation. Thirty ninegrams of SBE-β-CD were added to approximately 70 ml sodium phosphate,monobasic buffer and dissolved with mixing at room temperature. To thissolution, 300mg of clopidogrel bisulfate was added and dissolved in thesolution with stirring. The pH of the solution adjusted as needed withsodium hydroxide to pH 5.5 and the solution was brought to a finalvolume of 100 ml by the addition of buffer.

EXAMPLE 3

An aqueous solution of clopidogrel bisulfate was prepared. Theformulation contained SBE7-β-CD [DS=6.6] (39% wt./vol). The procedurewas identical to that of Example 2 except 0.1M sodium phosphate,monobasic buffer was used instead of a 0.2 M. The final pH was 5.52.

EXAMPLE 4

An aqueous solution of clopidogrel bisulfate was prepared at pH 7.97.The formulation comprised Captisol® (SBE-β-CD, DS=6.6) (39.0% wt./vol.)and clopidogrel bisulfate. The amounts used are specified in the tablebelow.

Ingredients Amount Clopidogrel Bisulfate 300.0 mg (equivalent to 229.9mg clopidogrel) SBE-β-CD 39.0 g (anhydrous basis) Sodium Phosphate,monobasic/ 0.2M dibasic buffer

The following procedure was used to prepare the formulation. Thirty ninegrams of SBE-β-CD were added to approximately 70 ml of sodium phosphate,monobasic/dibasic buffer and dissolved with mixing at room temperature.To this solution 300 mg of clopidogrel bisulfate was added and dissolvedin the solution with stirring. The pH of the solution was adjusted asneeded with sodium hydroxide to pH 7.97 and the solution was brought toa final volume of 100 ml with buffer.

EXAMPLE 5

An aqueous solution of clopidogrel bisulfate was prepared. Theformulation contained SBE-β-CD [DS=6.6] (39% wt./vol.). The procedurewas identical to that of Example 4 except 0.1M sodium phosphate,monobasic/dibasic buffer was used instead of a 0.2 M buffer. The finalpH was 8.03.

EXAMPLE 6

An aqueous solution of clopidogrel bisulfate was prepared at pH 5.50.The formulation comprised HP-β-CD [DS=4.3] (−25.0% wt./vol.) andclopidogrel bisulfate. The amounts used are specified in the tablebelow.

Ingredients Amount Clopidogrel Bisulfate 300.0 mg (equivalent to 229.9mg clopidogrel) HP-β-CD 25.0 g (anhydrous basis) Sodium Phosphate,monobasic buffer 0.2M

The following procedure was used to prepare the formulation. Twenty fivegrams of HP-β-CD were added to approximately 70 ml of sodium phosphate,monobasic buffer and dissolved with mixing at room temperature. To thissolution, 300 mg of clopidogrel bisulfate was added and dissolved in thesolution with stirring. The pH of the solution was adjusted as neededwith sodium hydroxide to pH 5.50 and the solution was brought to a finalvolume of 100 ml with buffer.

EXAMPLE 7

An aqueous solution of clopidogrel bisulfate was prepared. Theformulation contained HP-β-CD [DS=4.3] (25% wt./vol). The procedure wasidentical to that of Example 6 except 0.1 M sodium phosphate, monobasicbuffer was used instead of the 0.2 M. The final pH was 5.52.

EXAMPLE 8

An aqueous solution of clopidogrel bisulfate was prepared at pH˜8.0. Theformulation comprised HP-β-CD [DS=4.3] (-25.0% wt./vol.) and clopidogrelbisulfate. The amounts used are specified in the table below.

Ingredients Amount Clopidogrel Bisulfate 300.0 mg (equivalent to 229.9mg clopidogrel) HP-β-CD 25.0 g (anhydrous basis) Sodium Phosphate,monobasic/ 0.2M dibasic buffer

The following procedure was used to prepare the formulation. Twenty fivegrams of HP-β-CD were added to approximately 70 ml of sodium phosphate,monobasic/dibasic buffer and dissolved with mixing at room temperature.To this solution, 300mg of clopidogrel bisulfate was added and dissolvedin the solution with stirring. The pH of the solution was adjusted asneeded with sodium hydroxide to pH of 7.97 and the solution was broughtto a final volume of 100 ml with buffer.

EXAMPLE 9

An aqueous solution containing clopidogrel bisulfate and HP-β-CD[DS=4.3] (25% wt./vol.) was prepared. The procedure was similar to thatof Example 8 except 0.1 M sodium phosphate, monobasic/dibasic buffer wasused instead of the 0.2 M buffer. The final pH was 8.05.

EXAMPLE 10

An aqueous solution of clopidogrel bisulfate was prepared at pH-5.50.The formulation comprised Captisol® (SBE7-β-CD, DS=6.6) (37.0% wt./vol.)and clopidogrel bisulfate. The amounts used are specified in the tablebelow.

Ingredients Amount Clopidogrel Bisulfate 2.45 g (equivalent to 1.88 g ofclopidogrel) SBE-β-CD 92.5 g (anhydrous basis) Citric Acid/SodiumCitrate Buffer 0.2M

The following procedure was used to prepare the formulation. SBE-β-CD(DS=6.6) (92.5 g) were added to approximately 200 ml of citricacid/sodium citrate buffer (0.2M) and dissolved with mixing at roomtemperature. To this solution, 2.45 g of clopidogrel bisulfate was addedand dissolved in the solution with stirring. The clear solution wasbrought to a final volume of 250 ml with buffer and a final pH of 5.50.

EXAMPLE 11

An aqueous solution of clopidogrel bisulfate was prepared. Theformulation contained SBE-β-CD [DS=6.6] (37% wt./vol.). The procedurewas identical to that of Example 10 except the solution was filled into10 ml serum vials with a 2.5 ml fill volume. The vials were transferredto a freeze dryer, e.g., FTS Systems' Dura Dry tray dryer attached to aDura Dry II MP Condenser Module, and lyophilized. The lyophiles werethen reconstituted using sterile water to give clear solutions.

EXAMPLE 12

An aqueous solution of clopidogrel bisulfate was prepared at pH˜5.50.The formulation comprised Captisol® (SBE-β-CD, DS=6.6) (37.0% wt./vol.)and clopidogrel bisulfate. The amounts used are specified in the tablebelow.

Ingredients Amount Clopidogrel Bisulfate 2.45 g (equivalent to 1.88 g ofclopidogrel) SBE-β-CD 92.5 g (anhydrous basis) Citric Acid/SodiumCitrate Buffer 0.1M

The following procedure was used to prepare the formulation. 92.5 g ofSBE-β-CD were added to approximately 200 ml of citric acid/sodiumcitrate buffer (0.1 M) and dissolved with mixing at room temperature. Tothis solution, 2.45 g of clopidogrel bisulfate was added and dissolvedin the solution with stirring. The clear solution was brought to a finalvolume of 250 ml with buffer and a pH of 5.48.

EXAMPLE 13

An aqueous solution of clopidogrel bisulfate was prepared. Theformulation contained SBE-β-CD (DS=6.6) (37% wt./vol). The procedure wasidentical to that of Example 12 except the solution was lyophilizedusing a freeze dryer e.g. FTS Systems' Dura Dry tray dryer attached to aDura Dry II MP Condenser Module. The lyophiles were reconstituted usingsterile water to give clear solutions.

EXAMPLE 14

An aqueous solution of clopidogrel bisulfate was prepared. Theformulation comprised Captisol® (SBE-β-CD, DS=6.6) at a concentration of20 % wt./vol, excess clopidogrel bisulfate and a surfactant, polysorbate80 (T ween 80®″* and polyethylene glycol-15-hydroxystearate (Solutol®̂.Samples of surfactant (T ween 80® and Solutol®̂ at differentconcentrations without any Captisol® (SBE-β-CD, DS=6.6) were alsoformulated at a pH of about 5.5. The samples were analyzed forclopidogrel content after equilibration by mixing at room temperature of−20-25° C. for at least 24 hours. The molar amounts added of eachcomponent and the resultant clopidogrel amounts assayed in solution arespecified in Tables 14a, 14b, 14c and 14d.

TABLE 14a Concentration of Concentration Concentration of Clopidogrel ofCaptisol Tween 80 Solubilized M (% w/v) M (% w/v) M mg/ml 0.92 (20%) 0.076 (10%) 0.029 9.22 0.92 (20%) 0.038 (5%) 0.078 25.1 0.92 (20%)0.015 (2%) 0.064 20.8 0.92 (20%) 0.008 (1%) 0.056 18.1 0.92 (20%)  0.004 (0.5%) 0.023 7.40 0.92 (20%) 0.00076 (0.1%)  0.007 2.30 0.92(20%)    0 (0%) 0.015 4.69

TABLE 14b Concentration of Concentration of Clopidogrel Concentration ofTween 80 Solubilized Captisol M M (% w/v) M mg/ml 0  0.076 (10%) 0.10132.6 0 0.038 (5%) 0.124 39.9 0 0.015 (2%) 0.050 16.0 0 0.008 (1%) 0.05615.2 0   0.004 (0.5%) 0.047 8.89 0 0.00076 (0.1%)  0.005 1.67 0    0(0%) 0.0003 0.11

TABLE 14c Concentration of Concentration Concentration of Clopidogrel ofCaptisol Solutol Solubilized M (% w/v) mg/ml (% w/v) M mg/ml 0.92 (20%)100 (10%) 0.071 29.8 0.92 (20%) 50 (5%) 0.067 28.0 0.92 (20%)   25(2.5%) 0.043 17.6 0.92 (20%) 10 (1%) 0.021 8.9 0.92 (20%)  0 (0%) 0.0154.69

TABLE 14d Concentration of Concentration of Clopidogrel Concentration ofSolutol Solubilized Captisol M mg/ml (% w/v) M mg/ml 0 100 (10%) 0.09530.7 0 50 (5%) 0.063 20.3 0   25 (2.5%) 0.029 9.34 0 10 (1%) 0.008 2.500  0 (0%) 0.015 4.69

EXAMPLE 15

A sweetened aqueous solution of clopidogrel bisulfate was prepared. Theformulation comprised Captisol® (SBE-β-CD, DS=6.6) (37.7% wt./vol.),clopidogrel bisulfate and mannitol. The amounts used are specified inthe table below.

Ingredients Amount Clopidogrel Bisulfate 100 mg (~76.0 mg ofclopidogrel) SBE-β-CD 3.77 g (anhydrous basis) Mannitol 1.00 g

The following procedure was used to prepare the formulation. 3.77 g ofSBE-β-CD were added to approximately 7 ml of sterile filtered water. Tothis solution 100 mg of clopidogrel bisulfate and 1.0 Og of mannitolwere added and dissolved in the solution by vortexing. The solution wasbrought to a final volume of 10 ml. Two milliliters of the solution wereplaced in a 10 ml sterile vial and lyophilized using a freeze dryer e.g.FTS Systems' Dura Stop tray dryer attached to a Dura Dry II MP CondenserModule. The lyophile was reconstituted using sterile filtered water togive a clear solution with a final pH of 1.80.

EXAMPLE 16

The formulation comprised Captisol® (SBE-β-CD, DS=6.6) (−37%),clopidogrel bisulfate and mannitol. The procedure was similar to that ofExample 15 except, 200 mg of clopidogrel bisulfate (152 mg clopidogrel)was used. The reconstituted lyophile was a clear solution with a pH of1.52.

EXAMPLE 17

An aqueous solution of clopidogrel bisulfate was prepared. Theformulation comprised Captisol® (SBE-β-CD, DS=6.6) (−37%), clopidogrelbisulfate and D-glucose. The procedure was similar to that of Example 15except, D-glucose was used instead of mannitol and the final pH afterreconstitution was 1.81. The reconstituted solution was clear.

EXAMPLE 18

The formulation comprised Captisol® (SBE-β-CD, DS=6.6) (−37%),clopidogrel bisulfate and D-glucose. The procedure was similar to thatof Example 17 except, 200 mg of clopidogrel bisulfate (152 mgclopidogrel) was used. The reconstituted lyophile was a clear solutionwith a pH of 1.82.

EXAMPLE 19

The stability of clopidogrel liquid formulations was determined afterexposure to stress by either heat or fluorescent light. The formulationscontained clopidogrel bisulfate and equimolar amounts of differentcyclodextrins or cyclodextrin derivatives. All formulations contained3.0 mg/ml clopidogrel bisulfate, equivalent to 2.29 mg/ml clopidogrelfree base and cyclodextrin at 0.18 M. The cyclodextrin formulations wereprepared by dissolving the appropriate amount of cyclodextrin in 70 mlof sodium phosphate monobasic/dibasic buffer (0.1 M and 0.2 M), addingthe clopidogrel, and mixing until all the clopidogrel was dissolved. Thesolutions were brought to a final volume of 100 ml with phosphate bufferthen passed through a 0.22 micron Millex-GV Durapore filter. Thesolutions were evaluated at both pH˜5.5 and pH˜8.0 and at twotemperatures, 60° C. and 40° C. Each of the solutions was analyzed forcontent of clopidogrel and presence of degradants by HPLC. Aliquots (1.5m1) of each solution were also placed in 1 dram glass vials withTeflon-lined screw-caps and stored exposed to high intensity fluorescentlight (−25 cm from a bank of Sylvania Cool White 15 watt lamps) for 9days. At the end of the 9 day storage period, each of the samples wereassayed by the HPLC method and the amounts of each of the maindegradants calculated as a percentage of clopidogrel peak area appearingin the chromatogram.

The samples were analyzed using a Perkin-Elmer HPLC system using aβ-cyclodextrin column 200×4.0 mm and a mobile phase containing 65%Methanol: 35% 0.1% triethyl ammonium acetate solution at pH˜4 flowing at0.7 ml/min. Detection was by UV absorption at 230 nm. Clopidogrelretention time was −18 minutes using this analytical system.

EXAMPLE 20

An aqueous solution of clopidogrel bisulfate was prepared. Theformulation comprised Captisol® (SBE-β-CD, DS=6.6) (19.0% wt./vol.) andclopidogrel bisulfate. The amounts used are specified in the tablebelow.

Ingredients Amount Clopidogrel Bisulfate 150.8 mg (equivalent to 115.6mg of clopidogrel) SBE-β-CD 9.52 g (anhydrous basis)

To prepare the formulation, 9.52 grams of SBE-β-CD were added toapproximately 30 ml of sterile filtered water and dissolved with mixingat room temperature. To this solution 150.8 mg of clopidogrel bisulfatewere added and dissolved in the solution with stirring. The solution wasbrought to a final volume of 50 ml and the pH was adjusted to 5.47. Thedensity of the solution was determined to be 1.011 g/cc. Aliquots of 2.5ml of the solution were placed in 10 ml Schott glass vials andfreeze-dried. The samples were then used for the in-vivo bleeding studydescribed in Example 21.

EXAMPLE 21

A bleeding time study was performed using mice as the model animal. Themice were administered clopidogrel-containing formulations either byinjection into the tail vein or by oral gavage of doses of 0.1 ml per 20gram mouse. This is equivalent to doses of 6.5 mg/kg clopidogrelbisulfate (equivalent to 5 mg/kg clopidogrel base). At selected timesafter dosing, a standardized transection of the tip (0.3 mm) of eachtail was performed then the mice were immediately suspended verticallywith the distal 2 cm of each tail immersed in a test tube containingsaline at 37° C. The time required for beginning a 15 second period ofbleeding cessation was then determined. A maximum cessation time of 180seconds was used.

The SBE-β-CD-Clopidogrel formulations from Example 20 were prepared byreconstituting each vial with 5.1 ml sterile water for injection(calculated using the density of solution and water) resulting insolutions containing 1.3 mg/ml clopidogrel bisulfate. The solutions wereagitated well until all the solid had dissolved. This solutionformulation was used for both intravenous and oral solution dosing.

PLAVIX® ®the commercially available drug product was delivered as anoral suspension in methylcellulose. One tablet was ground up andsuspended in 37.5 ml of 0.5% w/v methylcellulose to give a suspensioncontaining 1.3 mg/ml clopidogrel bisulfate. The PLAVIX® suspension wasdelivered orally.

Aspirin at 100 mg/kg was used as a positive control.

The sampling regimen is as shown in the table below.

# of Mice per Sample Timepoint Timepoints Aspirin Control 5 30 minSBE-β-CD - Clopidogrel (oral) 5 5, 10, 20, 30 min SBE-β-CD - Clopidogrel(IV) 5 5, 10, 20, 30 min PLAVIX ® marketed product 5 5, 10, 20, 30 min(Tablet) ground and suspended in methylcellulose

EXAMPLE 22

The competitive binding study was conducted using clopidogrel bisulphateand aspirin (ASA, acetyl salicylic acid). The work was conducted on anauto-titrator such that the pH was maintained at −5.5 using sodiumhydroxide. The required amount of SBE-β-CD and aspirin was firstsolubilized in 0.2 μm Millipore filtered water then transferred to theauto-titrator. To this solution, excess clopidogrel bisulfate was addedand allowed to equilibrate for 2 hours while stirring and maintainingthe pH˜5.5. The amounts of SBE-β-CD and aspirin are shown in the tablebelow.

SBE-β-CD Content Aspirin Content M (mg/ml) M (mg/ml) 0.0134 (29.1 mg/ml)0.042 (7.5 mg/ml) 0.0125 (272 mg/ml) 0.111 (20 mg/ml) 0.0123 (268 mg/ml)0.166 (30 mg/ml) 0.0161 (34.8 mg/ml) No Aspirin

After equilibration, the solutions were sampled and analyzed forclopidogrel content.

EXAMPLE 23

Dissolution of the SBE-β-CD-clopidogrel bisulfate based formulation wascompared to the marketed PLAVIX formulation. Two forms of theclopidogrel formulation were made.

In the first formulation, a preformed complex of Captisol andclopidogrel bisulfate was made by solubilizing SBE-β-CD (2.5 g) in 100ml of sterile filtered water. To this, clopidogrel bisulfate (980 mg)was added. The solution was then spray dried using a Buchi 190 minispray dryer.

The second formulation comprised a physical mixture of SBE-β-CD (2.5 g)and clopidogrel bisulfate (980 mg), where the two ingredients wereweighed out and thoroughly mixed together in the solid state just priorto tableting. Magnesium stearate was added to the formulations as alubricant prior to tableting.

Each tablet contained magnesium stearate (0.5% wt/wt), SBE-β-CD (250 mg)and clopidogrel bisulfate (98 mg, which is equivalent to 75 mg of thefree base clopidogrel). This correlates with the marketed productPLAVIX® which has 75 mg of free base clopidogrel.

A comparative dissolution analysis was done using a Vankel vk7000dissolution apparatus. The paddle method was employed spinning at 50 rpmand media temperature maintained at 37° C. The dissolution media was1000 ml of a citric acid/disodium phosphate buffer at pH˜5.4. Eachtablet was tested in triplicate, and the sampling times were 5, 15, 30,45 and 60 minutes with each sample being filtered through a 35 μmfilter. These samples were then assayed for clopidogrel bisulfatecontent.

EXAMPLE 24

An aqueous solution of clopidogrel bisulfate and tirofiban hydrochloridemonohydrate is prepared at pH˜5.5. The formulation comprised Captisol®(SBE-β-CD, DS=6.6) (39.0% wt./vol.), clopidogrel bisulfate and tirofibanhydrochloride monohydrate. The amounts used are specified in the tablebelow.

Ingredients Amount Clopidogrel Bisulfate 100 mg (equivalent to ~76.0 mgof clopidogrel) Tirofiban hydrochloride 0.449 mg (equivalent to 0.4 mgtirofiban) monohydrate SBE-β-CD 3.90 g (anhydrous basis)

The following procedure is use to prepare the formulation. SBE-β-CD(3.90 g) is added to sterile water (approximately 7 ml) and dissolvedwith mixing at room temperature. To this solution, clopidogrel bisulfate(100 mg) is added and dissolved in the solution with stirring. Thentirofiban hydrochloride monohydrate (0.449 mg) is dissolved in thesolution, and the pH is adjusted as needed to pH˜5.5 with sodiumhydroxide. The solution is brought to a final volume of 10.0 ml by theaddition of sterile water.

EXAMPLE 25

An aqueous solution of clopidogrel bisulfate and enoxaparin sodium isprepared at pH-5.5. The formulation comprises Captisol® (SBE-β-CD,DS=6.6) (39.0% wt./vol.), clopidogrel bisulfate and enoxaparin sodium.The amounts used are specified in the table below.

Ingredients Amount Clopidogrel Bisulfate 100 mg (equivalent to ~76.0 mgof clopidogrel) Enoxaparin Sodium 10.5 mg SBE-β-CD 3.90 g (anhydrousbasis)

The following procedure is used to prepare the formulation. SBE-β-CD(3.90 g) is added to sterile water (approximately 7 ml) and dissolvedwith mixing at room temperature. To this solution, clopidogrel bisulfate(100 mg) is added and dissolved in the solution with stirring. Then,enoxaprin sodium (10.5 mg) is dissolved, and the pH is adjusted asneeded to pH˜5.5 with sodium hydroxide or hydrochloric acid. Thesolution is brought to a final volume of 10.0 ml by the addition ofsterile water.

EXAMPLE 26

A fresh whole blood sample is collected from a subject and plateletaggregation is measured with the point-of-care MICROS cell counter (ABXDiagnostics) and the Plateletworks test platform (Helena Laboratories).The cell counter uses traditional electronic impedance cell countingprinciples where a reference platelet count is performed on 1 mL offresh whole blood in a Plateletworks tube containing K₃-EDTA as theanticoagulant. The sample is then passed through the cell counter andthe platelet count is determined. This process is repeated with a second1 mL sample of fresh whole blood in a Plateletworks tube containing bothcitrate and 20 μmol/L ADP (Adenosine-5′-Diphosphate, the agonist). Inthe presence of ADP, platelets associate and aggregate. As theaggregated platelets exceed the threshold limitations for platelet size,they are no longer counted as individual platelets. The ratio of theplatelet count between the agonist and reference tubes is calculated aspercent platelet aggregation.

EXAMPLE 27

A open-label, dose-escalation clinical trial was conducted in healthyadult volunteers to determine the platelet aggregation inhibitioneffects and pharmacokinetics of an aqueous liquid formulation of theinvention as follows. The data obtained from this study are detailed inFIGS. 10-14 and discussed in further detail above.

Objectives:

To assess the safety and tolerability of single ascending doses of PM103 I.V.

To determine the dose-response of PM 103 LV. for inhibition ofADP-induced, platelet aggregation.

To measure plasma concentrations of clopidogrel, clopidogrel carboxylicacid, and clopidogrel thiol in plasma.

Design:

An open-label, ascending dose escalation study of six planned singleintravenous doses of PM103 LV. (0.1 mg, 1.0 mg, 10 mg, 30 mg, 100 mg and300 mg, clopidogrel).

Treatments:

Treatment: PM103 LV. (clopidogrel bisulfate); 0.1, 1.0, 10, 30 mg, 100mg or 300 mg

Subjects:

72 healthy male or female volunteers (12 per cohort) 18 years of age andolder were enrolled to the following planned cohorts based on subjecttolerability:

Cohort 1: PM103 LV. 0.1 mg, single dose

Cohort 2: PM103 LV. 1.0 mg, single dose

Cohort 3: PM103 LV. 10 mg, single dose

Cohort 4: PM103 LV. 100 mg, single dose

Cohort 5: PM103 LV. 30 mg, single dose

Cohort 6: PM103 LV. 300 mg, single dose

Inclusion Criteria:

Be a healthy male or female 18 years of age and older. Women ofchildbearing potential must be using a medically acceptable form ofbirth control for the duration of the trial and must have a negativeserum pregnancy test at screening and upon check-in to the studyfacility.

Have a BMI within the range of 18-35 kg/m², inclusive.

Be able to communicate effectively with the study personnel.

Have no significant disease or abnormal laboratory values as determinedby medical history, physical examination or laboratory evaluations,conducted at the screening visit or on admission to the clinic.

Have a normal 12-lead electrocardiogram, without any clinicallysignificant abnormalities of rate, rhythm or conduction.

Be nonsmokers defined as not having smoked in the past 6 months prior todosing.

Be adequately informed of the nature and risks of the study and givewritten informed consent prior to receiving study medication.

Have a normal platelet count >125,000/μL

Procedures:

PLAVIX® Pre-screen:

Subjects who successfully meet the screening criteria were scheduled fora clopidogrel dose effectiveness screen. The subject received a singleoral dose of PLAVIX® (300 mg) and was monitored for safety andtolerability and had platelet aggregation assessments performed predose(Screening Baseline) and at 2 hours after dosing.

Main Study:

Cohorts 1, 2, 3, 4 and 6 included two Phases. Phase 1 consisted of twosubjects. Phase 2 was dosed no less than 24 hours after Phase 1(assuming no safety concerns) and consisted of ten subjects. Studyprocedures were the same for both Phases. Cohort 5 consisted of twelvesubjects and all subjects were dosed on the same day. The procedures forthis study were as follows:

After a minimum post screening washout period of 14 days (including nouse of aspirin, other non-steroidal anti-inflammatory drugs, or otherdrugs known to affect platelet function) each eligible subject returnedto the clinic on the evening of Day -1. On Day 1 subjects will receivePM 103 I.V.

At approximately 1 hour prior to dosing, an IV port was inserted intothe antecubital region of the arm used for dosing (arm #1). For 0.1 mg(Cohort 1), 1.0 mg (Cohort 2), 10 mg (Cohort 3), and 30 mg (Cohort 5),at Time=0, a syringe containing PM 103 I.V. was attached to the IV portand the study formulation was administered as a bolus push injection (IVpush). For the 100 mg dose (Cohort 4), at Time=0, a glass bottlecontaining PM 103 I.V. 100 mg in 40 mL was attached to the IV port andthe study drug was administered as an infusion over 4 minutes (10niL/min). For the 300 mg dose (Cohort 6), at Time=0, a glass bottlecontaining PM103 I.V. 300 mg in 120 mL was attached to the IV port andthe study drug was administered as an infusion over 8 minutes (15niL/min). Immediately after the study drug administration was completed,the LV port was flushed with 3 mL of NSS and remained in the arm untilthe port was removed 1 hour after dose initiation. Subjects weredischarged from the clinic on Day 2 following collection of the 24-hourblood sample. Subjects came back at the clinic approximately 36 hourspost-dose (on Day 2) for assessment of adverse events.

Dosing in Cohort 1 (0.1 mg), Cohort 2 (1.0 mg) and Cohort 3 (10 mg) wasseparated by at least 7 days to enable safety and effectivenessevaluations prior to proceeding to the next dose level. Cohort 4 (100mg) was dosed at least 14 days following Cohort 3. Cohort 5 (30 mg) wasdosed at least 7 days following Cohort 4. Cohort 6 (300 mg) was dosed atleast 7 days following Cohort 5.

Pharmacokinetics:

Plasma samples for determination of clopidogrel, clopidogrel carboxylicacid, and clopidogrel thiol concentrations were obtained from arm #2(the arm not used for dosing) at baseline (pre-dose), 1, 5, 10, 20 and30 minutes, and 1, 2, 3, 4, 6, 8, 12, and 24 hours after doseinitiation.

Pharmacokinetic variables were calculated for clopidogrel, clopidogrelcarboxylic acid, and clopidogrel thiol using standard, non-compartmentalmethods. Calculations were conducted using WinNonlin (version 5.0.1).Standard variables were expected to include Cmax, Tmax, and AUCT.Variables λz, T1/2, and AUC were computed whenever sufficient pointswerer available in the terminal portion of the concentration versus timecurve.

Pharmacodynamics:

Blood samples for determination of ADP-induced platelet aggregationinhibition using the impedance method (see Dyszkiewicz-Korpanty et al.above) were obtained from arm #2 (the arm not used for dosing) atbaseline (check-in, Day-1), 15 and 30 minutes, and 2, 5, and 24 hoursafter dose initiation.

Statistical Analysis:

All pharmacokinetic results were summarized by treatment group usingappropriate descriptive statistics. Dose proportionality was assessedusing a plot of mean AUC, AUCT, and Cmax by dose level.

The ADP-induced platelet aggregation inhibition results using theimpedance method were summarized by treatment group and time point.Maximum % platelet inhibition and area under the % platelet inhibitioncurve was calculated for each individual and summarized by treatmentgroup

Safety Evaluations:

All safety variables (including adverse events, vital signsmeasurements, clinical laboratory results, electrocardiogram results,and other safety variables) were listed by subject and domain. Theincidence of all adverse events (AEs), treatment-emergent adverseevents, and treatment-related adverse events were tabulated by MedDRA™preferred term, system organ class, and treatment group. All laboratoryresults, vital sign measurements, and other safety variables weresummarized using appropriate descriptive statistics. The incidence oftreatment emergent laboratory abnormalities were summarized and listedby laboratory test. No hypothesis testing were performed.

All references cited herein are incorporated by reference.

The above is a detailed description of particular embodiments of theinvention. It will be appreciated that, although specific embodiments ofthe invention have been described herein for purposes of illustration,various modifications may be made without departing from the spirit andscope of the invention. Accordingly, the invention is not limited exceptby the appended claims. All of the embodiments disclosed and claimedherein can be made and executed without undue experimentation in lightof the present disclosure.

What is claimed is:
 1. A method of reducing the required therapeuticdose in a responder subject in need of clopidogrel to achieve a targettherapeutic effect therein, the method comprising: parenterally orperorally administering to the subject a first therapeutically effectiveamount of clopidogrel in a pharmaceutical composition comprising SAE-CDand clopidogrel, wherein the first therapeutically effective amount isat least 1.25-fold, at least 1.5-fold, at least 2-fold, at least 3-fold,at least 4-fold, at least 5-fold, at least 7-fold or at least 10-foldsmaller than a second therapeutically effective amount, which is theamount of clopidogrel required to provide substantially the same targettherapeutic effect when clopidogrel is administered to the subjectperorally in a reference tablet composition comprising clopidogrel andexcluding SAE-CD.
 2. The method of claim 1, wherein the firsttherapeutically effective amount is at least 1.1-fold, at least1.2-fold, at least 1.25-fold, at least 1.5-fold, at least 2-fold, atleast 3-fold, at least 4-fold, at least 5-fold, at least 7-fold, atleast 8-fold, at least 10-fold smaller, at least 15-fold, at least20-fold, about 1.1 to about 20-fold, about 1.2-fold to about 15-fold,about 1.25-fold to about 10-fold, about 2-fold to about 10-fold, orabout 3 fold to about 8-fold smaller than a second therapeuticallyeffective amount, which is the amount of clopidogrel required to providesubstantially the same therapeutic effect when clopidogrel isadministered to the subject perorally in a reference solidpharmaceutical composition excluding SAE-CD.
 3. The method of claim 2,wherein the first therapeutically effective amount is no more than 900mg, 750 mg, 675 mg, 600 mg, 450 mg, 375 mg, 300 mg, 225 mg, 200 mg, 150mg, 100 mg, 75 mg, 50 mg, 40 mg, 30 mg, 25 mg, 20 mg, 15 mg, 12.5 mg, 10mg, 7.5 mg, 5 mg, 2 mg, 1 mg, 0.75 mg, or 0.1 mg of clopidogrel, or inthe range of 0.1 to 900 mg, 0.1 to 100 mg, 50 mg to 600 mg, 100 to 300mg, about 300 mg, 300 to 600 mg, 300 to 900 mg, 600 to 900 mg, 50 to 600mg, 75 to 600 mg, 150 to 600 mg, or 200 to 450 mg, and the secondtherapeutically effective amount is 300 mg to 900 mg for solid oraltablets.
 4. A method of converting a subject that is a non-responder, interms of oral administration of clopidogrel, to a responder, the methodcomprising parenterally administering a pharmaceutical compositioncomprising SAE-CD and clopidogrel to a subject in need thereof, therebyproviding a therapeutic response to clopidogrel in the subject.
 5. Amethod of treating sickle cell crisis, comprising administering atherapeutically effective amount of a pharmaceutical compositioncomprising SAE-CD and clopidogrel, in combination with a secondtherapeutic agent is selected from the group consisting of: folic acid,hydroxyurea, NSAIDs, antibiotics, iron chelators, bronchodilators,diuretics, anxiolytics, α-agonisst, hydralazine, pentoxifylline,diltiazem, gonatropin-releasing hormone analogs, diethylstilbestrol, andcombinations of the foregoing.
 6. The method of claim 5, wherein thesecond therapeutic agent is formulated in the pharmaceuticalcomposition.
 7. The method of claim 5, wherein the second therapeuticagent is administered separately from the pharmaceutical composition. 8.The method of claim 5 wherein the pharmaceutical composition and thesecond therapeutic agent are independently administered perorally orparenterally.
 9. The method of claim 8, wherein pharmaceuticalcomposition and the second therapeutic agent are both administeredperorally.
 10. The method of claim 8, wherein pharmaceutical compositionand the second therapeutic agent are both administered parenterally.